Caracterización molecular del mutante riso 1508 de cebada : modificaciones transcripcionales y posttranscripcionales en el desarrollo de la semilla

La semilla es el principal órgano reproductivo de las plantas espermatofitas, permitiendo la dispersión de las poblaciones y asegurando su supervivencia gracias a su tolerancia a la desecación y a su capacidad para germinar bajo condiciones ambientales óptimas. El rendimiento y valor económico de los cereales, que constituyen la primera cosecha mundial, depende, en buena medida, de la eficacia con que se acumulan en la semilla sustancias de reserva: proteínas, carbohidratos y lípidos. El principal carbohidrato acumulado en la semilla de cebada es el almidón y la fracción mayoritaria de proteínas es la de las prolaminas (solubles en etanol al 70%); estas proteínas tienen muy bajo contenido en lisina, un aminoácido esencial en la dieta de animales monogástricos. Con el fin de mejorar el valor nutricional de la semilla de cebada, se han obtenido diferentes mutantes con un mayor contenido en este aminoácido. Riso 1508 es un mutante de cebada rico en lisina cuya mutación lys3a, de efectos pleiotrópicos, segrega como un único gen mendeliano. Entre otros, presenta una reducción drástica de la expresión de algunos genes que codifican proteínas de reserva de tipo prolamina, en concreto, presenta reducida la expresión de los genes que codifican B-, C- y ϒ-Hordeínas y del inhibidor de tripsina CMe, pero no tiene alterada la expresión del gen que codifica las D-Hordeínas. Este último gen carece en su promotor del motivo GLM (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), que es reconocido por factores transcripcionales bZIP. En este trabajo, el mutante de cebada Riso 1508 se ha utilizado como herramienta para profundizar en el conocimiento de la regulación génica en semillas durante las fases de la maduración y la germinación. Para ello, en una primera aproximación, se llevó a cabo un análisis transcriptómico comparando el genotipo mutante con el silvestre durante la maduración de la semilla. Además de confirmar variaciones en los genes que codifican proteínas de reserva, este análisis indicó que también estaban afectados los genes relacionados con metabolismo de carbohidratos. Por ello se decidió caracterizar la familia multigénica de sacarosas sintasa (SUSy) en cebada. Se anotaron dos nuevos genes, HvSs3 y HvSs4, cuya expresión se comparó con la de los genes HvSs1 y HvSs2, previamente descritos en el laboratorio. La expresión de los cuatro genes en tejidos diferentes y su respuesta a estreses abióticos se analizó mediante RT-qPCR. HvSs1 y HvSs2 se expresaron preferencialmente durante el desarrollo del endospermo, y HvSs1 también fue un tránscrito abundante durante la germinación. HvSs1 se indujo en hojas en condiciones de anoxia y HvSs3 por estrés hídrico, y ambos genes se indujeron por tratamientos de frío. La localización subcelular de las cuatro isoformas no fue sólo citoplásmica, sino que también se localizaron en zonas próximas a retículo endoplásmico y en la cara interna de la membrana plasmática; además, se observó una co-localización de HvSS1 con el marcador de mitocondrias. Estos datos sugieren un papel distinto aunque parcialmente solapante de las cuatro Sacarosa Sintasas de cebada, descritas hasta la fecha. Las cinéticas de expresión de los genes que codifican los TFs más importantes implicados en la regulación génica durante el desarrollo del endospermo de cebada, se analizaron por RT-qPCR en ambos genotipos, demostrando que los TFs de la clase DOF aparecieron desregulados durante todo el proceso en Riso 1508 comparado con el cv. Bomi, aunque también se observaron diferencias significativas en algunos de los que codifican bZIPs. Estudios previos indicaban que el ortólogo de BLZ2 en maíz, O2, se regula post-traduccionalmente mediante un mecanismo de fosforilación/defosforilación reversible, y que la forma defosforilada es la fisiológicamente activa. En este trabajo se demostró que BLZ2 está sujeto a este tipo de regulación y que la proteín-fosfatasa HvPP2C2 está implicada en el proceso. La interacción de HvPP2C2 y BLZ2 tiene lugar en el núcleo celular únicamente en presencia de 100 μM ABA. En el mutante Riso 1508, BLZ2 se encuentra en un estado hiperfosforilado tanto durante la maduración como durante la germinación de la semilla, lo que dificultaría la unión de BLZ2 a las secuencias GLM en los promotores de los genes que codifican B-, C-,y ϒ- Hordeínas y CMe. Summary The seed is the main reproductive organ of spermatophyte plants allowing the spread of populations and ensuring their survival through its desiccation tolerance and because of their ability to germinate under optimum environmental conditions. Yield and economic value of cereal crops, that constitute the first world crop, depend largely on the efficiency with which they accumulate in the seed reserve substances: proteins, carbohydrates and lipids. The main carbohydrate accumulated in the barley seed is starch and the major protein fraction is that of prolamins (soluble in 70% ethanol); these proteins have a very low lysine content, an essential amino-acid for the diet of monogastric animals. In order to improve the nutritional value of the barley seed, different mutants have been obtained with a higher content of this amino-acid. Riso 1508 is one lysine-rich mutant whose mutation (lys3a) segregates as a single Mendelian gene with pleiotropic effects, such as a drastic reduction of genes encoding the trypsin inhibitor CMe and the B-, C-and ϒ-hordeins, but has not altered the expression of the gene encoding the D-hordeins. This latter gene lacks in its promotor the GLM motif (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), that is recognised by bZIP transcription factors In this work we have used the barley mutant Riso 1508 as a tool for better understanding gene regulation in seeds during the maturation and germination phases. To this aim, a transcriptomic analysis was performed comparing wild and mutant genotypes during seed maturation. Besides confirming variations in the expression of genes encoding reserve proteins, this analysis indicated that some genes related with carbohydrate metabolism were also affected. It was therefore decided to characterize the multigene family of sucrose synthases (SUSy) in barley. Two new genes were annotated, HvSs3 and HvSs4, and its expression was compared with that of genes HvSs1 and HvSs2, previously described in our laboratory. The expression of the four genes in different tissues and in response to abiotic stresses was analyzed by RTqPCR. HvSs1 and HvSs2 were preferentially expressed during the development of the endosperm, and the HvSs1 transcript was also abundant upon germination. HvSs1 was induced in leaves by anoxic conditions, HvSs3 by water stress, and both genes were induced by cold treatments. The subcellular localization of all four isoforms was not only cytoplasmic, but they could be found along the endoplasmic reticulum and at the inner side of the cell membrane; HvSS1, was also associated with the mitochondrial marker. These data suggest a distinct but partially overlapping roles for the barley sucrose synthases, described so far. The expression kinetics of the genes encoding the most important TFs involved in gene regulation during barley endosperm development was analyzed by RT-qPCR in both genotypes. These data show that the genes encoding DOF TFs were mis-regulated throughout the process in Riso 1508, although significant differences were also found among some of those encoding bZIPs. Previous studies indicated that the BLZ2 orthologue in maize, O2, was post-translationally regulated by reversible phosphorylation/dephosphorylation and that the dephosphorylated protein is the physiologically active form. In this work we demostrate that BLZ2 is under a similar regulation and that the proteinphosphatase HvPP2C2 is implicated in the process. The interaction between HvPP2C2 and BLZ2 takes place in the cell nucleus only in the presence of 100 μM ABA. In the Riso 1508 mutant, BLZ2 is found in a hyperphosphorylated state in the maturation phase and upon seed germination; because of this, the BLZ2 binding to the GLM promoter sequences of genes encoding B-, C- y ϒ- Hordeins and CMe would be decreased in the mutant.

c&d-learning implementation architecture: Adapting e-learning to developing countries

From the educational point of view, the most widespread method in developing countries is on-site education. Technical and economic resources cannot support conventional distance learning infrastructures and it is even worse for courses in universities. They usually suffer a lack of qualified faculty staff, especially in technical degrees. The literature suggest that e-learning is a suitable solution for this problem, but its methods are developed attending to educational necessities of the First World and cannot be applied directly to other contexts. The proposed methodology is a variant of traditional e-learning adapted to the needs of developing countries. E-learning for Cooperation and Development (c&d-learning) is oriented to be used for educational institutions without adequate technical or human resources. In this paper we describe the c&d-learning implementation architecture based on three main phases: hardware, communication and software; e.g. computer and technical equipping, internet accessing and e-learning platform adaptation. Proper adaptation of educational contents to c&d-learning is discussed and a real case of application in which the authors are involved is described: the Ngozi University at Burundi.

Desarrollo de la radio en el marco de las rivalidades político-militares de principios del siglo XX

A lo largo de este proyecto, se analizarán los comienzos de la radio en las primeras décadas del siglo XX, teniendo como objetivo describir el desarrollo científico-tecnológico y las rivalidades político-militares de las potencias europeas de Reino Unido y Alemania. En primer lugar se verán los precursores de la radio que existían anteriormente al surgimiento de la radiotelegrafía como la telegrafía óptica, la telegrafía eléctrica o el teléfono. Al igual que los primeros investigadores, como James Clerk Maxwell o Heinrich Rudolf Hertz, gracias a los cuales el origen de la radio fue posible. También se analizará el contexto histórico-político de Reino Unido y Alemania donde se muestran las características de las dos grandes potencias del momento, el Imperio Británico y el Imperio Alemán. Además, la cada vez mayor importancia de la telegrafía sin hilos, hará necesaria una organización internacional, dando lugar a las Conferencias Internacionales de Radiotelegrafía y que enfrentarán a estas dos potencias por defender sus intereses. Guillermo Marconi, por parte de Reino Unido, y Telefunken, por parte de Alemania, serán las principales marcas comerciales de radiotelegrafía, no solo en sus respectivos países, sino también en el resto del mundo. Entre ambas compañías se verán las rivalidades por la incesante lucha de patentes, el desarrollo tecnológico y el control de los mercados. También se tratará el enfrentamiento directo que se produjo en la Primera Guerra Mundial, donde la radio jugaría un papel fundamental. Por último, se analizarán los comienzos de la radiodifusión en la segunda década del siglo XX, donde la radio ya estaba consolidada como uno de los medios de comunicación más importante de la época, capaz de llegar a todos los rincones del planeta y perfectamente integrada en la sociedad del momento. ABSTRACT. Throughout this project, it will be analyzed the beginning of the radio in the first decades of the XX century, having as goal to describe the scientific-technological development and the political-militaries opponents of the European’s countries, United kingdom and Germany. Firstly, it will be shown the radio’s precursors before the origin of the wireless telegraphy, like: optical telegraphy, electric telegraphy or telephone. Moreover, it will show the first researchers like: Clerk Maxwell or Heinrich Hertz, thanks to them, the origin of the radio was possible. In addition, it will analyze the historical-political context of United Kingdom and Germany, where it is shown the main features of the Britain Empire and the German Empire. Moreover, the importance of the telegraphy without wire will make necessary and international organization, resulting the International Conferences of Wireless Telegraphy and it will put together these main countries in order to protect their interests. Guglielmo Marconi, by United Kingdom, and Telefunken, by Germany, they will be the main commercials brand of the wireless telegraphy, not only in their countries, but also in all over the world. Between both companies will be seen the main competitions for their patens, the technology development and the market’s control. Moreover, it will be dealt the direct conflict in the First World of War, where the radio has a fundamental role. Finally, it will be analyze the beginning of the wireless broadcasting in the second decade of the XX century, where the radio was established as one of the most important media of the age, able to arrive at every places of the world and it was perfectly integrated in the society of that moment.

University Networking in Developing Countries based on C4D-Learning

In recent years, the establishment of cooperation networks between universities is one of the most important trends in higher education all over the world. Well recognized local and international university networks have been implemented in most educational institutions. It is common to find associations of various prestigious universities collaborating in a high-­‐technology research project including a very specialized teaching as well. This is the most common cooperation networks among higher education institutions in developed countries. An increasingly common type of networking between developed and developing universities is related to cooperation for development. This is the case of many universities in Africa that are needed for external help in order to improve its capabilities. Numerous memorandums of understanding regarding first world institutions that collaborate with universities in developing countries describe contributions of eventual visiting professors, teaching material and courses. But probably there exist another type of more important, but less explored association, such as networking among developing universities. The new goal, in this case, is not only the excellence but also the mutual development.

Technology transfer of renewable energy for rural development: historical evolution and lessons of experience from the model WWP

Technology transfer (TT) in the area of renewable energy (RE) throughout history has been an important tool for rural development (RD). Initially, the TT has been conceptualized as the purchase or donation of machinery from first world countries - without any consideration of staff training and contextual conditions for the adaptation of technology to the needs of the country. Various researches have revealed the existence of different approaches to planning the TT of RE, demonstrating the high complexity of projects from the social and contextual dimension. This paper addresses the conceptual evolution of the TT of RE for RD, examining its different periods considered for three criteria: historical events occurred, the role of stakeholders and changing objectives for the TT of RE for RD. For the conceptual analysis of changes the model Working With People (WWP) is used for planning and project management of high social complexity in RD. The analysis defines the existence of four historical periods in the TT of RE and synthesizes the lessons of experience from the three dimensions (ethical-social, technical-entrepreneurial, and political-contextual) of the WWP model.

La transformación de la morada: una investigación sobre los invariantes arquitectónicos en la vivienda contemporánea

Le Corbusier denominó la nueva casa como la "machine à habiter / máquina de habitar", adaptándola a la revolución industrial que desde el inicio del siglo XX estaba cambiando el mundo. La casa, entendida como una máquina, sería un objeto útil, fabricado mediante el ensamblaje de piezas industrializadas al igual que un coche, un avión o un barco. Pero más adelante, en el prólogo del libro ‘Precisiones’, titulado ‘Prólogo americano’. Le Corbusier traslada sus ideas después del viaje a las Américas, escritas en el barco de regreso a Europa y en ellas encontramos la prueba más evidente de que algo está cambiando en él. En este nuevo enfoque contribuyen probablemente muchas cosas, tal vez algunas nunca lleguemos a saberlas, pero entre las que con toda probabilidad lo hacen de forma decisiva, están algunos de sus viajes. Norte de África, España o América del Sur. En los suburbios de Buenos Aires o en Río, llenos de casas hechas de plancha ondulada, sin corazón y sin alma, y que, a pesar de todo, tienen uno y otra; Le Corbusier recibe una lección de arquitectura que nunca olvidará y que le llevará a decir: “he visto una vivienda obrera de plancha ondulada, pero muy bien puesta, en la cual un rosal adornaba la puerta. Era todo un poema de los tiempos modernos” O bien “busco con verdadero afán esas casas que son casas de hombres y no casas de arquitectos". Al final de su vida, libre de un compromiso político o de una atadura excesivamente estricta con la construcción; sus propuestas son más orgánicas y no tan estructurales, basta ver la Cabanon que se construye para él y su mujer. En este trabajo; iniciado sobre la base de la conferencia que Martin Heidegger pronuncia el 5 de agosto de 1951 en Darmstadt, “Bauen Wohnen Denken”, apoyándose en el escrito de Santa Teresa de Cepeda (1515-1582), y guiados por sus siete Moradas del “Castillo Interior”, escrito del año 1577; se estudia la transformación de la casa y la invariancia de la morada. La casa posee uno de los mayores poderes de integración para los pensamientos, los recuerdos y los sueños del hombre. La casa en la vida del hombre, suplanta contingencias, multiplica sus consejos de continuidad. Sin ella el hombre sería un ser disperso. Es cuerpo y alma. Es el primer mundo del ser humano... Y siempre, en sus sueños, la casa es una gran cuna, como nos dice Bachelard. Ha habido una evidente evolución del concepto de vivienda, a lo largo del tiempo pero aún así todas las moradas siguen siendo reflejo de sus habitantes. La casa y la morada son dos cosas distintas. Cuando un hombre hace suya una casa, encuentra su morada. ABSTRACT Le Corbusier named the new house as the "machine à habiter / machine for living", adapting it to the industrial revolution that was changing the world from the beginning of the twentieth century. The house, resembled as a machine, was a useful object made by the assembly of industrial pieces like a car, an airplane or a boat. But later, after his trip to the Americas, and reflected at the preface, 'American Prologue', of his book called 'Clarifications', Le Corbusier evolved his ideas into a new way of thinking. Wrote on the boat back to Europe, the lines written in this book are the clear evidence that something has changed in him. We might never get to know all of the variables that influenced his new approach, but we can be certain that the trips to South America, North Africa and Spain played a pivotal role on the evolution of his thinking. In the suburbs of Buenos Aires or Rio, where he was exposed to roughly made houses, without any heart or soul, but nevertheless owned by men, Le Corbusier learnt an unforgettable lesson. Hence, he would retrospect: "I saw a corrugated iron worker house, but very well decorated with a rosebush garnishing the front door. It was a poem of the modern times" or "I deeply look forward for those houses that are people’s homes not architect’s ones". At the end of his life, free of political commitment or an overly strict attachment to the construction, his proposals are more organic and less structural, as represented by the Cabanon built for himself and his wife. In this paper, based on Martin’s Heidegger conference in Darmstadt “Bauen Wohnen Denken” (august 5th of 1951) and supported by Santa Teresa de Cepeda’s (1515-1582) letter and her Seven Mansions of the Interior Castle, transformation and invariance is studied. The house has one of the greatest powers of integration for the thoughts, memories and dreams of men. The house, in the life of an individual, overcome men contingencies and multiplies men advice for continuity. Without it, men would be a dispersed human being. The house is body and soul. It is the first world of the human being ... And always, in his dreams, “the house is a large cradle”, as said by Bachelard. There has been a clear evolution of the concept of housing over time. But after all, the house still remains as a reflection of its inhabitants. When a man makes a house his own, we can conclude, he has found what he can call his “morada”.

Chemistry Education around the World: The First-Year Experience

En este trabajo se analizan y discuten las similitudes en el primer año de enseñanza de la química a nivel universitario en diferentes países. Este aspecto es una base fundamental que deben considerar, tanto profesores como alumnos, en programas de intercambio.

A first approach to the optimization of Bogotá's TransMilenio BRT system

Bus rapid transit (BRT) systems are massive transport systems with medium/high capacity, high quality service and low infrastructure and operating costs. TransMilenio is Bogotá's most important mass transportation system and one of the biggest BRT systems in the world, although it only has completed its third construction phase out of a total of eight. In this paper we review the proposals in the literature to optimize BRT system operation, with a special emphasis on TransMilenio, and propose a mathematical model that adapts elements of the above proposals and incorporates novel elements accounting for the features of TransMilenio system.

Technical evolution of 3d modular construction from the nineteenth century to world war II

3D Modular construction is poorly known and scarcely published in technical literature. In spite of that there are an increasing number of manufacturers offering their products in different countries. This method has largely evolved from early examples such as the American Gold Rush prefabrication in the nineteenth century, the Sears precut homes or Voisin´s prototypes for modular homes, to the end of the first half of the twentieth century. In this period a non negligible number of attempts in 3D modular construction have been carried out, ranging from theoretical proposals to several hundred or thousand units produced. Selected examples of modular architecture will be analyses in order to illustrate its technical evolution, concerning materials, structure, transportation and on site assembly. Success and failure factors of the different systems will be discussed. Conclusions about building criteria shown in them and their applicability in current architecture will be drawn.

World Space Observatory-Ultraviolet: ISSIS, the imaging instrument

The Imaging and Slitless Spectroscopy Instrument (ISSIS) will be flown as part of the science instrumentation in the World Space Observatory-Ultraviolet (WSO-UV). ISSIS will be the first UV imager to operate in a high Earth orbit from a 2 m class space telescope. In this contribution, the science driving the ISSIS design and the main characteristics of this instrument are presented.

First 20 years of DNDC: Model evolution and GRAMP.

The DNDC (DeNitrification and DeComposition) model was first developed by Li et al. (1992) as a rain event-driven process-orientated simulation model for nitrous oxide, carbon dioxide and nitrogen gas emissions from the agricultural soils in the U.S. Over the last 20 years, the model has been modified and adapted by various research groups around the world to suit specific purposes and circumstances. The Global Research Alliance Modelling Platform (GRAMP) is a UK-led initiative for the establishment of a purposeful and credible web-based platform initially aimed at users of the DNDC model. With the aim of improving the predictions of soil C and N cycling in the context of climate change the objectives of GRAMP are to: 1) to document the existing versions of the DNDC model; 2) to create a family tree of the individual DNDC versions; 3) to provide information on model use and development; and 4) to identify strengths, weaknesses and potential improvements for the model.