Evaluación de la vulnerabilidad sísmica urbana basada en tipologías constructivas y disposición urbana de la edificación. Aplicación en la ciudad de Lorca, región de Murcia

Actualmente, diversos terremotos han puesto de manifiesto la importancia de planificar las ciudades y la gran influencia que tiene el comportamiento de los edificios como consecuencia de los resultados de pérdidas humanas y económicas. Ante la imposibilidad de evitar la ocurrencia de terremotos y de predecirlos con un margen pequeño de tiempo para tomar acciones a corto plazo, la reducción de la vulnerabilidad de los elementos expuestos es la medida más eficaz para prevenir los daños y para evitar el desastre. Existen varios estudios anteriores de Norman B. Green (1980), Teresa Guevara López (2009 y 2012) que recogen criterios ya generalizados dentro de la bibliografía sísmica y algunos aspectos procedentes de norma sísmicas precursoras en este campo (por ejemplo, las peruanas) para establecer inicialmente unos principios urbanístico-sísmicos. Además, varios proyectos relacionados con el riesgo sísmico, RisK-Ue (2003), SERAMAR (Lars Abrahamczyk et al., 2013) han desarrollado metodologías que clasifican la vulnerabilidad de los edificios teniendo en cuenta modificadores por comportamientos y configuraciones irregulares sísmicamente. El presente trabajo desarrolla una metodología empírica para identificar y caracterizar los parámetros urbanísticos que determinan una respuesta sísmica irregular de las edificaciones, graduar su relación con el daño tras un terremoto y poder así disminuir la vulnerabilidad sísmica de las ciudades. La metodología desarrollada en esta tesis doctoral se aplica en la ciudad de Lorca, Región de Murcia. Se realiza un trabajo de campo donde se clasifican los edificios según su tipología estructural y sus parámetros urbanísticos. A través de un estudio estadístico se analiza la correlación con el daño de las edificaciones tras el terremoto del 11 de mayo de 2011. Previamente se ha hecho una clasificación de los edificios según la clase de suelo en la que se encuentran según el Eurocódigo8 (Navarro et al, 2012). Por último, se aplica la metodología para obtener una estimación de la habitabilidad de los edificios en Lorca post sismo. Para esta clasificación se ha adoptado el criterio recogido en diversas recomendaciones internacionales, la mayoría de las cuales se basan en la documentación generada por el ATC- Applied Technology Council, distinguiendo entre edificios habitables (no daño-daño no estructural) y edificios no habitables (daño estructural). ABSTRACT Currently, various earthquakes have made clear first, the importance of city planning and secondly, the great influence that has the behaviour of buildings as a consequence of the results of human and economic losses. Faced with the impossibility of avoiding the occurrence of earthquakes and predicting its with a small margin of time to take action in the short term, the reduction of the vulnerability of exposed elements is the most effective measure to prevent damage and to prevent the disaster. There are several previous studies, Norman B. Green (1980), Teresa Guevara López (2009-2012) collecting criteria already widespread within the seismic bibliography and we can find some aspects from standard seismic precursor in this field (for example, the Peruvian) to initially establish urban - seismic principles. In addition, several projects related to seismic risk, RisK-EU (2003), SERAMAR (Lars Abrahamczyk et al., 2013) have developed methodologies that classify the vulnerability of buildings taking into account modifiers for behaviours and irregular configurations in seismical terms. This paper develops an empirical methodology to identify and characterize the irregular urban parameters seismically, graduate its relationship with the building damages after an earthquake and thus reduce the seismic vulnerability of cities. The methodology developed in this thesis applies in the city of Lorca, Region of Murcia. Fieldwork where buildings are classified according to their structural type and its urban performance parameters. Through a statistical study the correlation with damage of buildings is analyzed after the earthquake of May 11, 2011. Previously a classification of the buildings has been made according to the kind of soil according to the Eurocodigo 8 (Navarro et al, 2012). Finally, you get an estimate of the building habitability in Lorca. As a result, this classification adopted the criterion contained in various international recommendations, most of which are based on the documentation published by the ATC - Applied Technology Council, habitable buildings (not damage -damage non-structural) and non habitable buildings (structural damage).

Performance of buildings with masonry infill walls during the 2011 Lorca earthquake

On Wednesday 11th May 2011 at 6:47 pm (local time) a magnitude 5.1 Mw earthquake occurred 6 km northeast of Lorca with a depth of around 5 km. As a consequence of the shallow depth and the small epicentral distance, important damage was produced in several masonry constructions and even led to the collapse of one of them. Pieces of the facades of several buildings fell down onto the sidewalk, being one of the reasons for the killing of a total of 9 people. The objective of this paper is to describe and analyze the failure patterns observed in reinforced concrete frame buildings with masonry infill walls ranging from 3 to 8 floors in height. Structural as well as non-structural masonry walls suffered important damage that led to redistributions of forces causing in some cases the failure of columns. The importance of the interaction between the structural frames and the infill panels is analyzed by means of non-linear Finite Element Models. The resulting load levels are compared with the member capacities and the changes of the mechanical properties during the seismic event are described and discussed. In the light of the results obtained the observed failure patterns are explained. Some comments are stated concerning the adequacy of the numerical models that are usually used during the design phase for the seismic analysis.

Stem CO2 efflux and its contribution to ecosystem CO2 efflux decrease with drought in a Mediterranean forest stand

tThe rate of metabolic processes demanding energy in tree stems changes in relation with prevailing cli-matic conditions. Tree water availability can affect stem respiration through impacts on growth, phloemtransport or maintenance of diverse cellular processes, but little is known on this topic. Here we moni-tored seasonal changes in stem CO2efflux (Fs), radial growth, sap flow and non-structural carbohydrates intrees of Quercus ilex in a Mediterranean forest stand subjected since 2003 to either partial (33%) through-fall exclusion (E) or unchanged throughfall (C). Fsincreased exponentially during the day by an effectof temperature, although sap flow attenuated the increase in Fsduring the day time. Over the year, Fsalso increased exponentially with increasing temperatures, but Fscomputed at a standard temperatureof 15?C (F15s) varied by almost 4-fold among dates. F15swas the highest after periods of stem growth anddecreased as tree water availability decreased, similarly in C and E treatments. The decline in F15swas notlinked to a depletion of soluble sugars, which increased when water stress was higher. The proportionof ecosystem respiration attributed to the stems was highest following stem growth (23.3%) and lowestduring the peak of drought (6.5%). High within-year variability in F15smakes unadvisable to pool annualdata of Fsvs. temperature to model Fsat short time scales (hours to months) in Mediterranean-type for-est ecosystems. We demonstrate that water availability is an important factor governing stem CO2effluxand suggest that trees in Mediterranean environments acclimate to seasonal drought by reducing stemrespiration. Stem respiratory rates do not seem to change after a long-term increase in drought intensity,however.

Assessment of expected damage on buildings subjected to Lorca earthquake through an energy-based seismic index method and nonlinear dynamic response analyses

The city of Lorca (Spain) was hit on May 11th, 2011, by two consecutive earth-quakes of magnitudes 4.6 and 5.2 Mw, causing casualties and important damage in buildings. Many of the damaged structures were reinforced concrete frames with wide beams. This study quantifies the expected level of damage on this structural type in the case of the Lorca earth-quake by means of a seismic index Iv that compares the energy input by the earthquake with the energy absorption/dissipation capacity of the structure. The prototype frames investigated represent structures designed in two time periods (1994–2002 and 2003–2008), in which the applicable codes were different. The influence of the masonry infill walls and the proneness of the frames to concentrate damage in a given story were further investigated through nonlinear dynamic response analyses. It is found that (1) the seismic index method predicts levels of damage that range from moderate/severe to complete collapse; this prediction is consistent with the observed damage; (2) the presence of masonry infill walls makes the structure very prone to damage concentration and reduces the overall seismic capacity of the building; and (3) a proper hierarchy of strength between beams and columns that guarantees the formation of a strong column-weak beam mechanism (as prescribed by seismic codes), as well as the adoption of counter-measures to avoid the negative interaction between non-structural infill walls and the main frame, would have reduced the level of damage from Iv=1 (collapse) to about Iv=0.5 (moderate/severe damage)

Comportamiento a fatiga de uniones a solape simple de acero prepintado y galvanizado con adhesivo silano modificado

(SPA) Se ha estudiado el comportamiento a fatiga de uniones adhesivas estructurales a solape simple. Los elementos estructurales fabricados en acero recubierto (galvanizado o pintado) sujetos a fluctuaciones de tensiones y deformaciones pueden fallar después de un número elevado de ciclos. El adhesivo silano modificado (MS) ha sido utilizado para la evaluación de la resistencia mecánica y a la fatiga de uniones de chapa delgada de acero protegido. El objetivo de este trabajo es analizar la influencia del recubrimiento en el comportamiento a fatiga de las uniones adhesivas con silano modificado. Las mejores propiedades se obtuvieron para el acero galvanizado con fallo cohesivo. Los resultados muestran que la compatibilidad química del adhesivo y la capa galvanizada mejoran la adhesión de la unión y la resistencia frente a cargas estáticas y dinámicas. Se ha obtenido el límite de fatiga para las uniones estudiadas. (ENG) The fatigue behaviour of single overlap of structural bonded joints was evaluated. Structural components made of thin coated steel (galvanised or painted) subjected to fluctuating stresses and strains may failure after a sufficient number of fluctuations. The modified silane adhesive (MS) has been used to evaluate the static and fatigue resistance of thin coated steel sheet joints. The aim of this paper is analyse the influence of coated on the fatigue behaviour of modified silane bonded joints. The best properties were obtained for galvanised steel with cohesive failure. The results show that chemical compatibility of adhesive and galvanized layer improve adhesion of joints and the mechanical resistance against static and dynamic loads. The fatigue limited has been obtained for joints studied.

Estudio de una metodología para transformar cualquier geometría en antifunicular añadiendo cargas externas

El antifunicular de cargas viene siendo empleado en la búsqueda de la mejor directriz para el diseño de arcos. Sin embargo, en muchas ocasiones, su forma acaba impuesta por condicionantes no meramente estructurales y su comportamiento dista de ser el óptimo. En este trabajo se propone un nuevo enfoque en el que son las cargas las que se adaptan a la geometría. Mediante la aplicación de una serie de cargas puntuales adicionales es posible obtener un comportamiento muy próximo a la compresión simple. Lo que se traduce en un importante descenso en las tensiones máximas que implica enormes ventajas. Las cargas se materializan mediante un cable de pretensado y se transmiten al arco por medio de barras, que pueden trabajar tanto a tracción como a compresión, dependiendo de la posición del cable con respecto a la estructura. Para sacar provecho a esta metodología, se ha realizado un programa que obtiene las diferentes configuraciones de este sistema para cualquier geometría introducida. Esta metodología permite diseñar estructuras de formas muy dispares dando solución a las demandas externas de una manera eficiente, elegante e innovadora. The antifunicular geometry has been employed as the optimal shape of a curved structure because it exhibits no bending under design loading. However, this shape is often defined by non-structural issues. Inevitably its behaviour is far from the ideal. The new approach, proposed in this study, consists in adding external loads to the main geometry in order to convert it into an antifunicular one. Adding point loads is possible to transform any geometry into a compression-only structure. This provides an important descent in terms of maximum stresses with great advantages for designing material-efficient structures. In the proposed system the loads are transmitted from a prestressed cable to the arch through struts, which can work in compression or in tension. It is possible to obtain different layouts of the cable for each starting shape. A computer programme has been implemented to solve it easily. This methodology allows to design structures of very different shapes, providing a solution to external demands in an efficient, innovative and elegant way.

Madera termo-tratada de frondosas para uso estructural

La madera termotratada es madera modificada mediante un proceso térmico a elevadas temperaturas que le proporciona mayor estabilidad dimensional y durabilidad sin incorporar productos químicos perjudiciales para el medio ambiente. Hasta el momento se ha aplicado fundamentalmente a madera de coniferas por motivos económicos, siendo su uso más habitual en ambientes exteriores o de elevada humedad, como elementos de revestimiento no estructurales, carpinterías, mobiliario de jardín, etc. En la presente tesis se estudia la viabilidad de la madera termotratada de frondosas para uso estructural, en particular fresno (Fraxinus excelsior L) y haya (Fagus sylvatica L). Con este fin, y considerando que el termotratamiento modifica la estructura interna de la madera resultando en un nuevo material, se realizan estudios experimentales y numéricos para su caracterización. Estos trabajos se desarrollan bajo el enfoque de la Mecánica de Fractura debido a la pérdida de resistencia y aumento de fragilidad que presenta el material, especialmente a tracción perpendicular a las fibras. Así mismo, se lleva a cabo una recopilación de las bases, fundamentos y metodologías de esta teoría aplicados a madera sin tratar y otros materiales debido a la inexistencia de este tipo de estudios en madera termotratada. De igual manera se realiza un programa de caracterización mecánica del material para determinar sus propiedades elásticas considerando un modelo ortótropo, necesarios en la investigación del comportamiento a fractura. El trabajo derivó en el desarrollo de un nuevo método de ensayo para la determinación multiparamétrica a partir de un sólo espécimen, proporcionando resultados mucho más robustos que los obtenidos con la metodología convencional de ensayos. En base a los trabajos realizados, considerando las limitaciones de resistencia y fragilidad, así como la dudosa aplicabilidad de las normativas existentes en madera sin tratar, se aconseja no utilizar tratamientos térmicos intensos en elementos estructurales primarios. Se propone su aplicación en elementos secundarios, de manera que un posible colapso no implique una pérdida de fiabilidad global de la estructura. Se estudia la viabilidad de un panel sandwich innovador y ecológico para fachadas expuesto a cargas de viento, compuesto de madera termotratada en las caras y panel aislante de fibras de madera con función estructural en el alma. Esta investigación se desarrolló dentro del proyecto de investigación Europeo "Holiwood", Holistic implementation of European thermal treated hardwood (TMT) in the sector of construction industry and noise protection by sustainable, knowledge-based and value added products, perteneciente al sexto Programa Marco. ABSTRACT Hcat-trcatcd wood is modified wood by a thermal process at high temperatures which provides greater dimensional stability and durability without adding harmful chemicals to the environment. It has been mainly applied to softwoods due mainly to economical reasons, being its most common use outdoors or in high humidity environments, as non-structural elements, furniture, etc. The present Thesis studies the feasibility of heat-treated hardwoods for structural uses, particularly ash (Fraxinus excelsior L) and beech (Fagus sylvatica L). To this end, and considering that heat treatment modifies the internal structure of the wood resulting in a new material, experimental and numerical studies are performed for its characterization. This investigation is developed under the approach of Fracture Mechanics due to the loss of strength and the increase in brittlcncss of the material, especially in tension perpendicular to the grain. Likewise, it holds a collection of the bases, foundations and methodologies of this theory applied to untreated wood and other materials due to the lack of such studies in heat-treated wood. In addition, studies for the mechanical characterization of the material are performed in order to determine the elastic properties considering an orthotropic model. This work is necessary in the investigation of the fracture behavior. It led to the development of a new test method for multiparameter determination by using just a single specimen, providing much more robust results than those obtained with conventional test methodology. Based on this investigation, and considering the limitations of strength and brittleness, and the questionable applicability of existing standards for untreated wood, it is advised not to use intense heat treatments in primary structural elements. It is proposed the application to secondary elements, so that a possible collapse does not involve a loss of overall reliability of the structure. It is studied the feasibility of an innovative and ecological sandwich panel for facades exposed to wind loads, composed by heat-treated wood faces and insulating wood fiberboard with structural function in the core. This investigación was developed within the European research project "Holiwood", Holistic implementation of European thermal treated hardwood (TMT) in the sector of construction industry and noise protection by sustainable, knowledge-based and value added products, of the Sixth Framework Program.

Functional response of Ulmus minor Mill. to drought, flooding and dutch elm disease

Ulmus minor es una especie arbórea originaria de Europa cuyas poblaciones han sido diezmadas por el hongo patógeno causante de la enfermedad de la grafiosis. La conservación de los olmos exige plantearse su propagación a través de plantaciones y conocer mejor su ecología y biología. Ulmus minor es un árbol de ribera, pero frecuentemente se encuentra alejado del cauce de arroyos y ríos, donde la capa freática sufre fuertes oscilaciones. Por ello, nuestra hipótesis general es que esta especie es moderadamente resistente tanto a la inundación como a la sequía. El principal objetivo de esta tesis doctoral es entender desde un punto de vista funcional la respuesta de U. minor a la inundación, la sequía y la infección por O. novo-ulmi; los factores que posiblemente más influyen en la distribución actual de U. minor. Con este objetivo se persigue dar continuidad a los esfuerzos de conservación de esta especie que desde hace años se dedican en varios centros de investigación a nivel mundial, ya que, entender mejor los mecanismos que contribuyen a la resistencia de U. minor ante la inoculación con O. novo-ulmi y factores de estrés abiótico ayudará en la selección y propagación de genotipos resistentes a la grafiosis. Se han planteado tres experimentos en este sentido. Primero, se ha comparado la tolerancia de brinzales de U. minor y U. laevis – otro olmo ibérico – a una inmersión controlada con el fin de evaluar su tolerancia a la inundación y comprender los mecanismos de aclimatación. Segundo, se ha comparado la tolerancia de brinzales de U. minor y Quercus ilex – una especie típica de ambientes Mediterránea secos – a la falta de agua en el suelo con el fin de evaluar el grado de tolerancia y los mecanismos de aclimatación a la sequía. El hecho de comparar dos especies contrastadas responde al interés en entender mejor cuales son los procesos que conducen a la muerte de una planta en condiciones de sequía – asunto sobre el que hay una interesante discusión desde hace algunos años. En tercer lugar, con el fin de entender mejor la resistencia de algunos genotipos de U. minor a la grafiosis, se han estudiado las diferencias fisiológicas y químicas constitutivas e inducidas por O. novo-ulmi entre clones de U. minor seleccionados a priori por su variable grado de resistencia a esta enfermedad. En el primer experimento se observó que los brinzales de U. minor sobrevivieron 60 días inmersos en una piscina con agua no estancada hasta una altura de 2-3 cm por encima del cuello de la raíz. A los 60 días, los brinzales de U. laevis se sacaron de la piscina y, a lo largo de las siguientes semanas, fueron capaces de recuperar las funciones fisiológicas que habían sido alteradas anteriormente. La conductividad hidráulica de las raíces y la tasa de asimilación de CO2 neta disminuyeron en ambas especies. Por el contrario, la tasa de respiración de hojas, tallos y raíces aumentó en las primeras semanas de la inundación, posiblemente en relación al aumento de energía necesario para desarrollar mecanismos de aclimatación a la inundación, como la hipertrofia de las lenticelas que se observó en ambas especies. Por ello, el desequilibrio del balance de carbono de la planta podría ser un factor relevante en la mortalidad de las plantas ante inundaciones prolongadas. Las plantas de U. minor (cultivadas en envases de 16 litros a media sombra) sobrevivieron por un prolongado periodo de tiempo en verano sin riego; la mitad de las plantas murieron tras 90 días sin riego. El cierre de los estomas y la pérdida de hojas contribuyeron a ralentizar las pérdidas de agua y tolerar la sequía en U. minor. Las obvias diferencias en tolerancia a la sequía con respecto a Q. ilex se reflejaron en la distinta capacidad para ralentizar la aparición del estrés hídrico tras dejar de regar y para transportar agua en condiciones de elevada tensión en el xilema. Más relevante es que las plantas con evidentes síntomas de decaimiento previo a su muerte exhibieron pérdidas de conductividad hidráulica en las raíces del 80% en ambas especies, mientras que las reservas de carbohidratos apenas variaron y lo hicieron de forma desigual en ambas especies. Árboles de U. minor de 5 y 6 años de edad (plantados en eras con riego mantenido) exhibieron una respuesta a la inoculación con O. novo-ulmi consistente con ensayos previos de resistencia. La conductividad hidráulica del tallo, el potencial hídrico foliar y la tasa de asimilación de CO2 neta disminuyeron significativamente en relación a árboles inoculados con agua, pero solo en los clones susceptibles. Este hecho enlaza con el perfil químico “más defensivo” de los clones resistentes, es decir, con los mayores niveles de suberina, ácidos grasos y compuestos fenólicos en estos clones que en los susceptibles. Ello podría restringir la propagación del hongo en el árbol y preservar el comportamiento fisiológico de los clones resistentes al inocularlos con el patógeno. Los datos indican una respuesta fisiológica común de U. minor a la inundación, la sequía y la infección por O. novo-ulmi: pérdida de conductividad hidráulica, estrés hídrico y pérdida de ganancia neta de carbono. Pese a ello, U. minor desarrolla varios mecanismos que le confieren una capacidad moderada para vivir en suelos temporalmente anegados o secos. Por otro lado, el perfil químico es un factor relevante en la resistencia de ciertos genotipos a la grafiosis. Futuros estudios deberían examinar como este perfil químico y la resistencia a la grafiosis se ven alteradas por el estrés abiótico. ABSTRACT Ulmus minor is a native European elm species whose populations have been decimated by the Dutch elm disease (DED). An active conservation of this species requires large-scale plantations and a better understanding of its biology and ecology. U. minor generally grows close to water channels. However, of the Iberian riparian tree species, U. minor is the one that spread farther away from rivers and streams. For these reasons, we hypothesize that this species is moderately tolerant to both flooding and drought stresses. The main aim of the present PhD thesis is to better understand the functional response of U. minor to the abiotic stresses – flooding and drought – and the biotic stress – DED – that can be most influential on its distribution. The overarching goal is to aid in the conservation of this emblematic species through a better understanding of the mechanisms that contribute to resistance to abiotic and biotic stresses; an information that can help in the selection of resistant genotypes and their expansion in large-scale plantations. To this end, three experiments were set up. First, we compared the tolerance to experimental immersion between seedlings of U. minor and U. laevis – another European riparian elm species – in order to assess their degree of tolerance and understand the mechanisms of acclimation to this stress. Second, we investigated the tolerance to drought of U. minor seedlings in comparison with Quercus ilex (an oak species typical of dry Mediterranean habitats). Besides assessing and understanding U. minor tolerance to drought at the seedling stage, the aim was to shed light into the functional alterations that trigger drought-induced plant mortality – a matter of controversy in the last years. Third, we studied constitutive and induced physiological and biochemical differences among clones of variable DED resistance, before and following inoculation with Ophiostoma novo-ulmi. The goal is to shed light into the factors of DED resistance that is evident in some genotypes of U. minor, but not others. Potted seedlings of U. minor survived for 60 days immersed in a pool with running water to approximately 2-3 cm above the stem collar. By this time, U. minor seedlings died, whereas U. laevis seedlings moved out of the pool were able to recover most physiological functions that had been altered by flooding. For example, root hydraulic conductivity and leaf photosynthetic CO2 uptake decreased in both species; while respiration initially increased with flooding in leaves, stems and roots possibly to respond to energy demands associated to mechanisms of acclimation to soil oxygen deficiency; as example, a remarkable hypertrophy of lenticels was soon observed in flooded seedlings of both species. Therefore, the inability to maintain a positive carbon balance somehow compromises seedling survival under flooding, earlier in U. minor than U. laevis, partly explaining their differential habitats. Potted seedlings of U. minor survived for a remarkable long time without irrigation – half of plants dying only after 90 days of no irrigation in conditions of high vapour pressure deficit typical of summer. Some mechanisms that contributed to tolerate drought were leaf shedding and stomata closure, which reduced water loss and the risk of xylem cavitation. Obviously, U. minor was less tolerant to drought than Q. ilex, differences in drought tolerance resulting mostly from the distinct capacity to postpone water stress and conduct water under high xylem tension among species. More relevant was that plants of both species exhibited similar symptoms of root hydraulic failure (i.e. approximately 80% loss of hydraulic conductivity), but a slight and variable depletion of non-structural carbohydrate reserves preceding dieback. Five- and six-year-old trees of U. minor (planted in the field with supplementary watering) belonging to clones of contrasted susceptibility to DED exhibited a different physiological response to inoculation with O. novo-ulmi. Stem hydraulic conductivity, leaf water potential and photosynthetic CO2 uptake decreased significantly relative to control trees inoculated with water only in DED susceptible clones. This is consistent with the “more defensive” chemical profile observed in resistant clones, i.e. with higher levels of saturated hydrocarbons (suberin and fatty acids) and phenolic compounds than in susceptible clones. These compounds could restrict the spread of O. novo-ulmi and contribute to preserving the near-normal physiological function of resistant trees when exposed to the pathogen. These results evidence common physiological responses of U. minor to flooding, drought and pathogen infection leading to xylem water disruption, leaf water stress and reduced net carbon gain. Still, seedlings of U. minor develop various mechanisms of acclimation to abiotic stresses that can play a role in surviving moderate periods of flood and drought. The chemical profile appears to be an important factor for the resistance of some genotypes of U. minor to DED. How abiotic stresses such as flooding and drought affect the capacity of resistant U. minor clones to face O. novo-ulmi is a key question that must be contemplated in future research.

Unearthing the roots of degradation of Quercus pyrenaica coppices: an integrative perspective from clonal structure to carbon budgets

Quercus pyrenaica es una especie rebrotadora de raíz intensa e históricamente aprovechada en monte bajo para la obtención de leñas, carbón y pastos. Debido al éxodo rural y a la aparición de nuevas fuentes energéticas, este aprovechamiento fue abandonado en la década de 1970. Desde entonces, las bajas producciones de madera y bellota y el puntisecado de los pies evidencian el generalizado estancamiento de estas masas. Uno de los mayores retos actuales de la selvicultura en el ámbito mediterráneo es encontrar usos alternativos para estos montes abandonados, siendo la conversión a monte alto una de las alternativas preferidas. Se han realizado resalveos de conversión, sin embrago, éstos se aplican sin un conocimiento integral de las causas de la degradación. En esta tesis doctoral, estudiamos un hipotético desequilibrio entre la parte radical y la parte aérea (R:S) de las cepas de rebollo como causa subyacente de su decaimiento. En una parcela experimental, aprovechada al menos desde el siglo XII, se realizaron análisis genéticos a priori para elucidar la estructura genética del rodal, y así estudiar la influencia del tamaño clonal en el funcionamiento de las cepas. Las cepas de mayor tamaño presentaron un menor crecimiento diametral de sus pies, así como mayores tasas de respiración radical, estimadas a partir de flujos internos de CO2 a través del xilema (FT) y de los flujos de CO2 del suelo. Estos resultados sugieren que el desequilibrio R:S aumenta con el tamaño clonal, dado que la eliminación periódica de órganos aéreos, al mismo tiempo que las raíces permanecen intactas, da lugar a un gran desarrollo del sistema radical que consume gran parte de los carbohidratos no estructurales (NSC) en respiración de mantenimiento, comprometiendo así el desarrollo de órganos aéreos. Se excavaron y pesaron dos cepas compuestas por cuatro y ocho pies, las cuales mostraron ratios R:S (0.5 y 1, respectivamente) superiores a los registrados en pies de origen sexual. Al igual que en otras especies rebrotadoras de raíz, se observaron altas concentraciones de NSC en las raíces (> 20% en primavera) y una gran proporción de albura en el sistema radical (52%) que alberga una notable reserva de NSC (87 kg en la cepa de mayor tamaño). En el sistema radical de dicha cepa, estimada mediante dataciones radiocarbónicas en 550 años de edad, se contaron 248 uniones radicales. La persistencia de sistemas radicales grandes, viejos, y altamente interconectados sugiere que la gran cantidad de recursos almacenados y consumidos en las raíces compensan un pobre desarrollo aéreo con una alta resiliencia vegetativa. Para un mejor entendimiento de los balances de carbono y del agotamiento de NSC en las cepas de rebollo, se midieron los flujos internos y externos de CO2 en troncos y los flujos de CO2 del suelo, y se estimó la respiración de órganos aéreos (RS) y subterráneos (RR). Estacionalmente, RS y RR reflejaron las dinámicas de flujo de savia y de crecimiento del tronco, y estuvieron determinadas principalmente por los flujos externos de CO2, dada la escasa contribución de FT a RS y RR (< 10% y < 2%, respectivamente). En una escala circadiana, la contribución de FT a RS aumentó hasta un 25% en momentos de alta transpiración. Las bajas concentraciones de CO2 en el xilema ([CO2] hasta un 0.11%) determinaron comparativamente unos bajos FT, probablemente causados por una limitada respiración del xilema y una baja resistencia a la difusión radial del CO2 impuestos por la sequía estival. Los pulsos de [CO2] observados tras las primeras lluvias de otoño apoyan esta idea. A lo largo del periodo vegetativo, el flujo medio de CO2 procedente del suelo (39 mol CO2 day-1) fue el mayor flujo respiratorio, tres y cuatro veces superior a RS (12 mol CO2 day-1) y RR (8-9 mol CO2 day-1), respectivamente. Ratios RR/RS menores que la unidad evidencian un importante peso de la respiración aérea como sumidero de carbono adicional. Finalmente, se ensayó el zanjado de raíces y el anillamiento de troncos como tratamientos selvícolas alternativos con el objetivo de aumentar las reservas de NSC en los troncos de las cepas. Los resultados preliminares desaconsejan el zanjado de raíces por el alto coste derivado posiblemente de la cicatrización de las heridas. El anillado de troncos imposibilitó el transporte de NSC a las raíces y aumentó la concentración de almidón por encima de la zona anillada, mientras que sistema radical se mantiene por los pies no anillados de la cepa. Son necesarias más mediciones y datos adicionales para comprobar el mantenimiento de esta respuesta positiva a largo plazo. Para concluir, destacamos la necesidad de estudios multidisciplinares que permitan una comprensión integral de la degradación de los rebollares ibéricos para poder aplicar a posteriori una gestión adecuada en estos montes bajos abandonados. ABSTRACT Quercus pyrenaica is a vigorous root-resprouting species intensively and historically coppiced for firewood, charcoal and woody pastures. Due to the rural exodus and the appearance of new energy sources, coppicing was abandoned towards 1970. Since then, tree overaging has resulted in stand stagnation displayed by slow stem growth, branch dieback, and scarce acorn production. The urgent need to find new alternative uses for abandoned coppices is recognized as one of the biggest challenges which currently faces Mediterranean silviculture; conversion into high forest by thinning is one of the preferred alternatives. For this aim, thinning has been broadly applied and seldom tested, although without a comprehensive understanding of the causes of stand stagnation. In this PhD study, we test the hypothesis of an imbalance between above- and below-ground organs, result of long term coppicing, as the underlying cause of Q. pyrenaica decay. In an experimental plot coppiced since at least the 12th century, genetic analyses were performed a priori to elucidate inconspicuous clonal structure of Q. pyrenaica to evaluate how clonal size affects the functioning of these multi-stemmed trees. Clonal size negatively affected diametric stem growth, whereas root respiration rates, measured by internal fluxes of CO2 through xylem (FT) and soil CO2 efflux, increased with clonal size. These results suggest root-to-shoot (R:S) imbalance intensifying with clonal size: periodic removal of aboveground organs whilst belowground organs remain undisturbed may have led to massive root systems which consume a great proportion of non-structural carbohydrates (NSC) for maintenance respiration, thus constraining aboveground performance. Furthermore, excavation of two multi-stemmed trees, composed by four and eight stems, revealed R:S ratios (0.5 and 1, respectively) greater than those reported for sexually regenerated trees. Moreover, as similarly observed in several root-resprouting species, NSC allocation to roots was favored ([NSC] > 20% in spring): a large proportion of sapwood maintained throughout the root system (52%) stored a remarkable NSC pool of 87 kg in the case of the largest clone. In this root system of the eight-stemmed tree, 248 root connections were counted and, by radiocarbon dating, its age was estimated to be 550-years-old. Persistence of massive, old and highly interconnected root systems suggests that enhanced belowground NSC storage and consumption reflects a trade-off between vegetative resilience and aboveground development. For a better understanding of tree carbon budget and the potential role of carbon starvation in Q. pyrenaica decay, internal and external stem CO2 fluxes and soil CO2 effluxes were monitored to evaluate respiratory costs above- and below-ground. On a seasonal scale, stem and root respiration (RS and RR) mirrored sap flow and stem growth dynamics. Respiration was determined to the greatest extent by external fluxes of CO2 to the atmosphere or soil, since FT accounted for a low proportion of RS and RR (< 10% and < 2%, respectively). On a diel scale, the contribution of FT to RS increased up to 25% at high transpiration rates. Comparatively low FT was determined by the low concentration of xylem CO2 registered ([CO2] as low as 0.11%), likely as a consequence of constrained xylem respiration and reduced resistance to CO2 radial diffusion imposed by summer drought. Xylem [CO2] pulses following first autumn rains support this idea. Averaged over the growing season, soil CO2 efflux was the greatest respiratory flux (39 mol CO2 day-1), three and four times greater than RS (12 mol CO2 day-1) and RR (8-9 mol CO2 day-1), respectively. Ratios of RR/RS below one evidence an additional and important weight of aboveground respiration as a tree carbon sink. Finally, root trenching and stem girdling were tested as complimentary treatments to thinning as a means to improve carbon reserves in stems of clonal trees. Preliminary results discouraged root trenching due to the high cost likely incurred for wound closure. Stem girdling successfully blocked NSC translocation downward, increasing starch concentrations above the girdled zone whilst the root system is fed by non-girdled stems within the clone. Further measurements and ancillary data are necessary to verify that this positive effect hold over time. To conclude, the need of multidisciplinary approaches for an integrative understanding on the functioning of abandoned Q pyrenaica coppices is highlighted for an appropriate management of these stands.

The making of the architecture of the plant cell wall: How cells exploit geometry

Cell wall deposition is a key process in the formation, growth, and differentiation of plant cells. The most important structural components of the wall are long cellulose microfibrils, which are synthesized by synthases embedded in the plasma membrane. A fundamental question is how the microfibrils become oriented during deposition at the plasma membrane. The current textbook explanation for the orientation mechanism is a guidance system mediated by cortical microtubules. However, too many contraindications are known in secondary cell walls for this to be a universal mechanism, particularly in the case of helicoidal arrangements, which occur in many situations. An additional construction mechanism involves liquid crystalline self-assembly [A. C. Neville (1993) Biology of Fibrous Composites: Development Beyond the Cell Membrane (Cambridge Univ. Press, Cambridge, U.K.)], but the required amount of bulk material that is able to equilibrate thermally is not normally present at any stage of the wall deposition process. Therefore, we have asked whether the complex ordered texture of helicoidal cell walls can be formed in the absence of direct cellular guidance mechanisms. We propose that they can be formed by a mechanism that is based on geometrical considerations. It explains the genesis of the complicated helicoidal texture and shows that the cell has intrinsic, versatile tools for creating a variety of textures. A compelling feature of the model is that local rules generate global order, a typical phenomenon of life.

NMR characterization of altered lignins extracted from tobacco plants down-regulated for lignification enzymes cinnamylalcohol dehydrogenase and cinnamoyl-CoA reductase

Homologous antisense constructs were used to down-regulate tobacco cinnamyl-alcohol dehydrogenase (CAD; EC 1.1.1.195) and cinnamoyl-CoA reductase (CCR; EC 1.2.1.44) activities in the lignin monomer biosynthetic pathway. CCR converts activated cinnamic acids (hydroxycinnamoyl–SCoAs) to cinnamaldehydes; cinnamaldehydes are then reduced to cinnamyl alcohols by CAD. The transformations caused the incorporation of nontraditional components into the extractable tobacco lignins, as evidenced by NMR. Isolated lignin of antisense-CAD tobacco contained fewer coniferyl and sinapyl alcohol-derived units that were compensated for by elevated levels of benzaldehydes and cinnamaldehydes. Products from radical coupling of cinnamaldehydes, particularly sinapaldehyde, which were barely discernible in normal tobacco, were major components of the antisense-CAD tobacco lignin. Lignin content was reduced in antisense-CCR tobacco, which displayed a markedly reduced vigor. That lignin contained fewer coniferyl alcohol-derived units and significant levels of tyramine ferulate. Tyramine ferulate is a sink for the anticipated build-up of feruloyl–SCoA, and may be up-regulated in response to a deficit of coniferyl alcohol. Although it is not yet clear whether the modified lignins are true structural components of the cell wall, the findings provide further indications of the metabolic plasticity of plant lignification. An ability to produce lignin from alternative monomers would open new avenues for manipulation of lignin by genetic biotechnologies.

Functional domains for assembly of histones H3 and H4 into the chromatin of Xenopus embryos

Histones H3 and H4 have a well defined structural role in the nucleosome and an established role in the regulation of transcription. We have made use of a microinjection strategy using Xenopus embryos to define the minimal structural components of H3 and H4 necessary for nucleosome assembly into metazoan chromosomes in vivo. We find that both the N-terminal tail of H4, including all sites of acetylation, and the C-terminal α-helix of the H4 histone fold domain are dispensable for chromatin assembly. The N-terminal tail and an N-terminal α-helix of H3 are also dispensable for chromatin assembly. However, the remainder of the H3 and H4 histone folds are essential for incorporation of these proteins into chromatin. We suggest that elements of the histone fold domain maintain both nucleosomal integrity and have distinct functions essential for cell viability.

Modulation of Ca2+ entry by polypeptides of the inositol 1,4,5-trisphosphate receptor (IP3R) that bind transient receptor potential (TRP): Evidence for roles of TRP and IP3R in store depletion-activated Ca2+ entry

Homologues of Drosophilia transient receptor potential (TRP) have been proposed to be unitary subunits of plasma membrane ion channels that are activated as a consequence of active or passive depletion of Ca2+ stores. In agreement with this hypothesis, cells expressing TRPs display novel Ca2+-permeable cation channels that can be activated by the inositol 1,4,5-trisphosphate receptor (IP3R) protein. Expression of TRPs alters cells in many ways, including up-regulation of IP3Rs not coded for by TRP genes, and proof that TRP forms channels of these and other cells is still missing. Here, we document physical interaction of TRP and IP3R by coimmunoprecipitation and glutathione S-transferase-pulldown experiments and identify two regions of IP3R, F2q and F2g, that interact with one region of TRP, C7. These interacting regions were expressed in cells with an unmodified complement of TRPs and IP3Rs to study their effect on agonist- as well as store depletion-induced Ca2+ entry and to test for a role of their respective binding partners in Ca2+ entry. C7 and an F2q-containing fragment of IP3R decreased both forms of Ca2+ entry. In contrast, F2g enhanced the two forms of Ca2+ entry. We conclude that store depletion-activated Ca2+ entry occurs through channels that have TRPs as one of their normal structural components, and that these channels are directly activated by IP3Rs. IP3Rs, therefore, have the dual role of releasing Ca2+ from stores and activating Ca2+ influx in response to either increasing IP3 or decreasing luminal Ca2+.

Endocytic Clathrin-coated Pit Formation Is Independent of Receptor Internalization Signal Levels

The mechanisms responsible for coated pit formation in cells remain unknown, but indirect evidence has argued both for and against a critical role of receptor cytoplasmic domains in the process. If the endocytic motifs of receptors are responsible for recruiting AP2 to the plasma membrane, thereby driving coated pit formation, then the level of constitutively internalized receptors at the membrane would be expected to govern the steady-state level of coated pits in cells. Here we directly test this hypothesis for broad classes of receptors containing three distinct constitutive internalization signals. Chimeric proteins consisting of an integral membrane reporter protein (Tac) coupled to cytoplasmic domains bearing tyrosine-, di-leucine-, or acidic cluster/casein kinase II-based internalization signals were overexpressed to levels that saturated the internalization pathway. Quantitative confocal immunofluorescence microscopy indicated that the number of plasma membrane clathrin-coated pits and the concentration of their structural components were invariant when comparing cells expressing saturating levels of the chimeric receptors to nonexpressing cells or to cells expressing only the Tac reporter lacking cytoplasmic internalization signals. Biochemical analysis showed that the distribution of coat proteins between assembled coated pits and soluble pools was also not altered by receptor overexpression. Finally, the cellular localizations of AP2 and AP1 were similarly unaffected. These results provide a clear indication that receptor endocytic signals do not determine coated pit levels by directly recruiting AP2 molecules. Rather, the findings support a model in which coated pit formation proceeds through recruitment and activation of AP2, likely through a limited number of regulated docking sites that act independently of endocytic signals.

An Intronic Enhancer Is Required for Deflagellation-induced Transcriptional Regulation of a Chlamydomonas reinhardtii Dynein Gene

Chlamydomonas reinhardtii flagellar regeneration is accompanied by rapid induction of genes encoding a large set of flagellar structural components and provides a model system to study coordinate gene regulation and organelle assembly. After deflagellation, the abundance of a 70-kDa flagellar dynein intermediate chain (IC70, encoded by ODA6) mRNA increases approximately fourfold within 40 min and returns to predeflagellation levels by ∼90 min. We show by nuclear run-on that this increase results, in part, from increased rates of transcription. To localize cis induction elements, we created an IC70 minigene and measured accumulation, in C. reinhardtii, of transcripts from the endogenous gene and from introduced promoter deletion constructs. Clones containing 416 base pairs (bp) of 5′- and 2 kilobases (kb) of 3′-flanking region retained all sequences necessary for a normal pattern of mRNA abundance change after deflagellation. Extensive 5′- and 3′- flanking region deletions, which removed multiple copies of a proposed deflagellation-response element (the tub box), did not eliminate induction, and the IC70 5′-flanking region alone did not confer deflagellation responsiveness to a promoterless arylsulfatase (ARS) gene. Instead, an intron in the IC70 gene 5′-untranslated region was found to contain the deflagellation response element. These results suggest that the tub box does not play an essential role in deflagellation-induced transcriptional regulation of this dynein gene.