Cartografía de QTL asociados a la tolerancia a estrés hídrico en frijol común (Phaseolus vulgaris L.)

Phaseolus vulgaris L. (frijol común o judía) es una leguminosa de gran demanda para la nutrición humana y un producto agrícola muy importante. Sin embargo, la producción de frijol se ve limitada por presiones ambientales como la sequía. En México, el 85% de la cosecha de frijol se produce en la temporada de primavera-verano, principalmente en las regiones del altiplano semiárido con una precipitación anual entre 250 y 400 mm. A pesar del implemento de tecnología en el campo, los factores naturales impiden al agricultor llegar a los rendimientos deseados. El Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP), como instituto de investigación gubernamental en México, tiene como objetivo la mejora de cultivos estratégicos, uno de ellos, P. vulgaris. Los estudios en relación a la sequía se enfocan especialmente en la selección de genotipos tolerantes, los cuales son sometidos en condiciones de estrés y monitoreando parámetros como el rendimiento y peso de semilla, además de algunos indicadores tales como índice de cosecha. El resultado de estos trabajos ha sido la obtención de variedades con mayor tolerancia a la sequía, tales como Pinto Villa y Pinto Saltillo. En los últimos años se ha avanzado notablemente en el conocimiento de las bases moleculares en las respuestas de las plantas al estrés. De acuerdo a diversos estudios se ha demostrado que las plantas bajo estrés por sequía experimentan cambios en la expresión de genes involucrados en la señalización, regulación de la transcripción y la traducción, transporte de agua y la función directa en la protección celular. También se ha observado que el déficit de agua es causado por las temperaturas extremas y la alta concentración de sales, por lo que al nivel molecular, las respuestas al estrés tienen puntos de especificidad y puntos de entrecruzamiento. La sequía puede generar estreses secundarios, tales como el nutricional, oxidativo y osmótico. Sin embargo, es necesario identificar y caracterizar muchos de los componentes involucrados en las respuestas al déficit hídrico, la caracterización de estos genes permitirá tener una mejor comprensión de los mecanismos bioquímicos y fisiológicos involucrados en la tolerancia al estrés. Actualmente, con el apoyo de la biología molecular se han identificado algunos genes que otorgan ventajas para la adaptación a ambientes desfavorables. Por lo que el objetivo del presente trabajo es identificar marcadores genéticos asociados a rasgos fenotípicos con énfasis a la tolerancia a estrés hídrico en P. vulgaris. Una vez establecidos los marcadores asociados al estrés hídrico, es factible considerar su uso para la selección asistida por marcadores en líneas o variedades de frijol de interés para los mejoradores. Se evaluaron 282 familias F3:5 derivadas de la cruza entre los cultivares Pinto Villa y Pinto Saltillo. Las familias se sembraron bajo un diseño simple de látice 17x17, el experimento se llevo acabo en el ciclo primavera-verano del 2010 y 2011, y otoñoinvierno de 2010 en el Campo Experimental Bajío del INIFAP con dos repeticiones para cada tratamiento de humedad (riego completo y sequía terminal). En todos los genotipos se realizó el fenotipado (variables fenotípicas) y el genotipado a través de marcadores moleculares. Los análisis estadísticos se basaron en el análisis de componentes principales (Eigen Analysis Selection Index Method, ESIM), la asociación entre marcadores SNP y el fenotipado (paquete SNPassoc para R) y el análisis de varianza (ANOVA). Los valores ESIM mostraron que las variables de Rendimiento, Días a floración, Días a madurez fisiológica e Índice de cosecha fueron sobresalientes en sequía terminal, por lo que se sugieren tomarse en consideración para los estudios de sequía en P. vulgaris como monitores de evaluación a la resistencia. Se identificaron nueve familias sobresalieron por sus valores ESIM (PV/PS6, 22, 131, 137, 149, 154, 201, 236 y 273), además de presentar valores superiores para el rendimiento en comparación con los parentales. Estos genotipos son candidatos interesantes para realizar estudios de identificación de loci asociados con la respuesta al estrés, y como potenciales parentales en el desarrollo de nuevas variedades de frijol. En los análisis de asociación SNPassoc se identificaron 83 SNPs significativos (p<0,0003) asociados a los rasgos fenotípicos, obteniendo un total de 222 asociaciones, de las cuales predomina el modelo genético de codominancia para las variables Días a floración, Periodo reproductivo y Biomasa total. Treinta y siete SNPs se identificaron a diferentes funciones biológicas a través del análisis de anotación funcional, de los cuales 12 SNPs (9, 18, 28, 39, 61, 69, 80, 106, 115, 128, 136 y 142) sobresalen por su asociación al fenotipado, y cuya anotación funcional indica que se encuentran en genes relacionados a la tolerancia a la sequía, tales como la actividad kinasa, actividad metabólica del almidón, carbohidratos y prolina, respuesta al estrés oxidativo, así como en los genes LEA y posibles factores de transcripción. En el caso de los análisis ANOVA, se identificaron 72 asociaciones entre los SNPs y las variables fenotípicas (F< 3,94E-04). Las 72 asociaciones corresponden a 30 SNPs y 7 variables fenotípicas, de las que predomina Peso de 100 semillas y Periodo reproductivo. Para los rasgos de Rendimiento, Índice de cosecha y Días a madurez fisiológica se presentaron asociaciones con seis SNPs (17, 34, 37, 50, 93 y 107), de los cuales, a los SNP37 y SNP107 fueron identificados a la anotación biológica de protein binding. Por otro lado, los SNP106 y SNP128 asociados al Periodo reproductivo, son genes con actividad kinasa y actividad metabólica del almidón, respectivamente. Para los marcadores tipo AFLP, se identificaron 271 asociaciones (F<2,34E-04). Las asociaciones corresponden a 86 AFLPs con todas las variables fenotípicas evaluadas, de las que predomina peso de 100 semillas, Días a floración y Periodo reproductivo. Debido a que los en los AFLPs no es posible determinar su anotación biológica, se proponen como marcadores potenciales relacionados a la resistencia a la sequía en frijol. Los AFLPs candidatos requieren más estudios tales como la secuenciación de los alelos respectivos, así como la identificación de éstas secuencias en el genoma de referencia y su anotación biológica, entre otros análisis, de esta manera podríamos establecer aquellos marcadores candidatos a la validación para la selección asistida. El presente trabajo propone tanto genotipos como marcadores genéticos, que deben ser validados para ser utilizados en el programa de mejoramiento de P. vulgaris, con el objetivo de desarrollar nuevas líneas o variedades tolerantes a la sequía. ABSTRACT Phaseolus vulgaris L. (common bean or judia) is a legume of great demand for human consumption and an important agricultural product. However, the common bean production is limited by environmental stresses, such as drought. In Mexico, 85% of the common bean crop is produced in the spring-summer season mainly in semiarid highland regions with a rainfall between 250 and 400 mm per year. In spite of the improvement of crop technology, the natural factors hamper getting an optimal yield. The National Institute for Forestry, Agriculture and Livestock (INIFAP) is a government research institute from Mexico, whose main objective is the genetic breeding of strategic crops, like P. vulgaris L. The drought tolerance studies particularly focus on the selection of bean tolerant genotypes, which are subjected to stress conditions, by means of monitoring parameters such as yield and seed weight, plus some agronomic indicators such as harvest index. The results of these works have led to obtain cultivars with higher drought tolerance such as Pinto Villa and Pinto Saltillo. Significant achievements have been recently made in understanding the molecular basis of stress plant responses. Several studies have shown that plants under drought stress present changes in gene expression related to cell signalling, transcriptional and translational regulation, water transport and cell protection. In addition, it has been observed that the extreme temperatures and high salt concentrations can cause a water deficiency so, at the molecular level, stress responses have specific and crossover points. The drought can cause secondary stresses, such as nutritional, oxidative and osmotic stress. It is required the identification of more components involved in the response to water deficit, the characterization of these genes will allow a better understanding of the biochemical and physiological mechanisms involved in stress tolerance. Currently, with the support of molecular biology techniques, some genes that confer an advantage for the crop adaptation to unfavourable environments have been identified. The objective of this study is to identify genetic markers associated with phenotypic traits with emphasis on water stress tolerance in P. vulgaris. The establishment of molecular markers linked to drought tolerance would make possible their use for marker-assisted selection in bean breeding programs. Two hundred and eighty two F3:5 families derived from a cross between the drought resistant cultivars Pinto Villa and Pinto Saltillo were evaluated. The families were sowed under a 17x17 simple lattice design. The experiment was conducted between spring-summer seasons in 2010 and 2011, and autumn-winter seasons in 2010 at the Bajio Experimental Station of INIFAP with two treatments (full irrigation and terminal drought). All families were phenotyped and genotyped using molecular markers. Statistical analysis was based on principal component analysis (Eigen Analysis Selection Index Method, ESIM), association analysis between SNP markers and phenotype (SNPassoc package R) and analysis of variance (ANOVA). The ESIM values showed that seed yield, days to flowering, days to physiological maturity and harvest index were outstanding traits in terminal drought treatment, so they could be considered as suitable parameters for drought-tolerance evaluation in P. vulgaris. Nine outstanding families for the ESIM values were identified (PV/PS6, 22, 131, 137, 149, 154, 201, 236 and 273), in addition, these families showed higher values for seed yield compared to the parental cultivars. These families are promising candidates for studies focused on the identification of loci associated to the stress response, and as potential parental cultivars for the development of new varieties of common bean. In the SNPassoc analysis, 83 SNPs were found significantly associated (p<0.0003) with phenotypic traits, obtaining a total of 222 associations, most of which involved the traits days to flowering, reproductive period and total biomass under a codominant genetic model. The functional annotation analysis showed 37 SNPs with different biological functions, 12 of them (9, 18, 28, 39, 61, 69, 80, 106, 115, 128, 136 and 142) stand out by their association to phenotype. The functional annotation suggested a connection with genes related to drought tolerance, such as kinase activity, starch, carbohydrates and proline metabolic processes, responses to oxidative stress, as well as LEA genes and putative transcription factors. In the ANOVA analysis, 72 associations between SNPs and phenotypic traits (F<3.94E- 04) were identified. All of these associations corresponded to 30 SNPs markers and seven phenotypic traits. Weight of 100 seeds and reproductive period were the traits with more associations. Seed yield, harvest index and days to physiological maturity were associated to six SNPs (17, 34, 37, 50, 93 and 107), the SNP37 and SNP107 were identified as located in protein binding genes. The SNP106 and SNP128 were associated with the reproductive period and belonged to genes with kinase activity and genes related to starch metabolic process, respectively. In the case of AFLP markers, 271 associations (F<2.34E-04) were identified. The associations involved 86 AFLPs and all phenotypic traits, being the most frequently associated weight of 100 seeds, days to flowering and reproductive period. Even though it is not possible to perform a functional annotation for AFLP markers, they are proposed as potential markers related to drought resistance in common bean. AFLPs candidates require additional studies such as the sequencing of the respective alleles, identification of these sequences in the reference genome and gene annotation, before their use in marker assisted selection. This work, although requires further validation, proposes both genotypes and genetic markers that could be used in breeding programs of P. vulgaris in order to develop new lines or cultivars with enhanced drought-tolerance.

Percolation in soils at Different Bulk Densities: Porosity, Connectivity and Grey Threshold.

To improve percolation modelling on soils the geometrical properties of the pore space must be understood; this includes porosity, particle and pore size distribution and connectivity of the pores. A study was conducted with a soil at different bulk densities based on 3D grey images acquired by X-ray computed tomography. The objective was to analyze the effect in percolation of aspects of pore network geometry and discuss the influence of the grey threshold applied to the images. A model based on random walk algorithms was applied to the images, combining five bulk densities with up to six threshold values per density. This allowed for a dynamical perspective of soil structure in relation to water transport through the inclusion of percolation speed in the analyses. To evaluate separately connectivity and isolate the effect of the grey threshold, a critical value of 35% of porosity was selected for every density. This value was the smallest at which total-percolation walks appeared for the all images of the same porosity and may represent a situation of percolation comparable among bulks densities. This criterion avoided an arbitrary decision in grey thresholds. Besides, a random matrix simulation at 35% of porosity with real images was used to test the existence of pore connectivity as a consequence of a non-random soil structure.

Respuestas anatómico fisiológicas frente a estrés hídrico en plantaciones de especies de crecimiento rápido para la producción de biomasa

El objetivo general de la presente Tesis es identificar algunas de las características anatómico-fisiológicas que confieren la capacidad de alcanzar una mejor productividad bajo clima mediterráneo a plantas de diversos genotipos de los géneros Populus y Eucalyptus, caracterizados por su carácter pionero, elevado crecimiento y vulnerabilidad a la cavitación. En los dos primeros capítulos se hace un seguimiento de la conductancia estomática a una selección de clones de eucalipto cultivados en invernadero, sometidos a diferentes dotaciones hídricas. Se realizaron además mediciones periódicas del pH de la savia del tallo y de la pérdida de conductividad hidráulica para investigar su implicación en la regulación química e hidraúlica del cierre estomático. Las variaciones en el pH de la savia obtenidas parecen responder a cambios en el déficit de presión de vapor de agua atmosférico y no a diferencias en la disponibilidad de agua en el suelo. La conductancia estomática presentó una correlación positiva significativa con el pH de la savia, pero no con la conductividad hidráulica. La variabilidad de la conductividad hidráulica máxima se discute a la luz de recientes investigaciones sobre los materiales constituyentes de las membranas de las punteaduras. Los clones que mostraron mayores conductancias hidráulicas y estomáticas presentaron valores más altos de producción y supervivencia, poniendo de manifiesto la utilidad del estudio de estas variables. Por el contrario, los valores más bajos de conductancia estomática e hidraúlica se encontraron en clones que han resultado un fracaso en plantaciones comerciales, en particular, fue destacable el mal resultado de un clon procedente de autocruzamiento respecto de otros híbridos. En el tercer capítulo de la tesis se estudian características anatómicas y funcionales del xilema relacionadas con la eficiencia en el transporte de agua a las hojas, y que pueden afectar directa o indirectamente a la transpiración y al crecimiento. Los estudios anatómicos fueron realizados sobre brotes anuales de chopo en una plantación situada en Granada, en condiciones de riego limitante. La combinación de rasgos anatómicos más favorable de cara a la producción de biomasa fue una densidad alta de vasos de diámetro intermedio. Los clones más productivos figuraron entre los más resistentes a la cavitación. Para estudiar el crecimiento de masas arboladas se utilizan frecuentemente parámetros fisiológicos como el índice de area foliar (LAI). La estimación del LAI a partir de fotografías hemisféricas aplicada a tallares de chopo plantados a alta densidad y a turno corto para biomasa se lleva a cabo mediante una metodología reciente empleada y discutida en el cuarto capítulo de la Tesis. Los resultados muestran que las diferencias de producción existentes entre genotipos, localidades de medición con diferentes dosis de riego, y años, pueden predecirse a partir de la determinación del índice de área foliar tanto por métodos directos como indirectos de estimación. Tanto los estudios realizados en eucalipto como en chopo han mostrado que los genotipos con menores producciones de biomasa en campo alcanzaron los menores valores de conductancia estomática en las condiciones más favorables así como el menor número de vasos en el xilema. La estrecha relación entre crecimiento y LAI confirma una vez más la importancia del desarrollo de la copa para sostener un buen crecimiento. El mayor desarrollo de la copa y rendimiento en biomasa se midieron en uno de los clones con un número de vasos más elevado, y menor vulnerabilidad a la cavitación en condiciones de estrés. Estos resultados ponen de manifiesto la importancia de las características anatómicas y funcionales del xilema como condicionantes del patrón de crecimiento de las plantas y el comportamiento de los estomas. ABSTRACT A number of anatomical xylem traits and physiological variables were analyzed in genotypes of both the Populus and Eucalyptus genera with the main aim of identifying traits in the genotypes which confer the ability to produce an acceptable biomass yield under Mediterranean climatic conditions. In the first two chapters of this PhD, the results of two experiments carried out on several clones of the species Eucalyptus globulus Labill. are presented. Chapters three and four include the results of another two trials on four poplar hybrid genotypes. One of the initial plant responses to water stress is stomatal closure, which can be triggered by hydraulic and/or chemical signals. The two first chapters of this PhD deal with trials in which stomatal conductance and percentage loss of hydraulic conductivity were monitored on a set of eucalyptus clones supplied by ENCE (former National Cellulose Company) and currently used in the company’s own commercial plantings. The experimental trials were carried out in greenhouses and the plants were submitted to two different watering regimes. The pH of the stem sap was periodically measured as the greenhouse temperature and humidity changed. The aim of these measurements was to investigate the role of both sap pH and percentage loss of hydraulic conductivity on stomatal regulation. The results obtained suggest that changes in sap pH are a response to vapor pressure deficit changes rather than to differences in soil water availability. We found significant correlation between stomatal conductance and sap pH, although no significant relationship was found between stomatal conductance and hydraulic conductivity. Variability in maximum hydraulic conductivity is discussed based on recent pit membrane constituent research. The study of hydraulic conductivity proved helpful in order to detect the clones with both higher growth and greater chance of survival, since clones displaying the lowest hydraulic conductivities were those that failed in commercial plantings. Anatomical xylem traits define the water transport efficiency to leaves and can therefore limit transpiration and growth. The third chapter of this PhD addresses anatomical xylem traits in poplar. One year old stem samples were taken from a water-stressed trial in Granada. The anatomical xylem study proved useful for detecting the lowest yielding genotypes. Clones with intermediate vessel size and high vessel densities were found to be those with the highest biomass yield. Differences in cavitation resistance depending on the clone tested and the water treatment applied were also found. The clones with the highest biomass yield were found to be among the most cavitation resistant clones in each watering regime. Xylem and physiological traits along with stomatal behavior are useful tools to determine plant growth. In order to study plantings or forests, it is more common to employ other physiological variables such as leaf area index (LAI). LAI estimation from hemispherical photographs applied to short rotation woody crops is a recently developed method that still requires fine tuning through further investigation. In the fourth chapter, data from LAI monitoring over two consecutive years were analyzed in two different locations where different irrigation treatments were applied. The results showed that differences in yield between genotypes, different irrigation regimes and years could be predicted by using the LAI estimates, either through direct or indirect estimation methods. Our studies of poplar and eucalyptus have shown that the field-grown genotypes with the lowest biomass yield displayed the lowest values of stomatal conductance under the most favorable environmental conditions and also had a low number of xylem conduits. The close relationship between LAI and growth highlights the importance of crown development in biomass growth. The highest LAI and biomass yield were recorded in one of the clones with higher vessel density and the lowest vulnerability to cavitation under stress conditions. These results underline the importance of research into anatomical and functional traits as factors influencing plant growth patterns and stomatal behavior.

Fractal Behaviour and Irregularity of Wetting Fronts in Heterogeneous Porous Media

The wetting front is the zone where water invades and advances into an initially dry porous material and it plays a crucial role in solute transport through the unsaturated zone. Water is an essential part of the physiological process of all plants. Through water, necessary minerals are moved from the roots to the parts of the plants that require them. Water moves chemicals from one part of the plant to another. It is also required for photosynthesis, for metabolism and for transpiration. The leaching of chemicals by wetting fronts is influenced by two major factors, namely: the irregularity of the fronts and heterogeneity in the distribution of chemicals, both of which have been described by using fractal techniques. Soil structure can significantly modify infiltration rates and flow pathways in soils. Relations between features of soil structure and features of infiltration could be elucidated from the velocities and the structure of wetting fronts. When rainwater falls onto soil, it doesn?t just pool on surfaces. Water ?or another fluid- acts differently on porous surfaces. If the surface is permeable (porous) it seeps down through layers of soil, filling that layer to capacity. Once that layer is filled, it moves down into the next layer. In sandy soil, water moves quickly, while it moves much slower through clay soil. The movement of water through soil layers is called the the wetting front. Our research concerns the motion of a liquid into an initially dry porous medium. Our work presents a theoretical framework for studying the physical interplay between a stationary wetting front of fractal dimension D with different porous materials. The aim was to model the mass geometry interplay by using the fractal dimension D of a stationary wetting front. The plane corresponding to the image is divided in several squares (the minimum correspond to the pixel size) of size length ". We acknowledge the help of Prof. M. García Velarde and the facilities offered by the Pluri-Disciplinary Institute of the Complutense University of Madrid. We also acknowledge the help of European Community under project Multi-scale complex fluid flows and interfacial phenomena (PITN-GA-2008-214919). Thanks are also due to ERCOFTAC (PELNoT, SIG 14)

Diagnosing Causes of Water Scarcity in Complex Water Resources Systems and Identifying Risk Management Actions

From the water management perspective, water scarcity is an unacceptable risk of facing water shortages to serve water demands in the near future. Water scarcity may be temporary and related to drought conditions or other accidental situation, or may be permanent and due to deeper causes such as excessive demand growth, lack of infrastructure for water storage or transport, or constraints in water management. Diagnosing the causes of water scarcity in complex water resources systems is a precondition to adopt effective drought risk management actions. In this paper we present four indices which have been developed to evaluate water scarcity. We propose a methodology for interpretation of index values that can lead to conclusions about the reliability and vulnerability of systems to water scarcity, as well as to diagnose their possible causes and to propose solutions. The described methodology was applied to the Ebro river basin, identifying existing and expected problems and possible solutions. System diagnostics, based exclusively on the analysis of index values, were compared with the known reality as perceived by system managers, validating the conclusions in all cases

Modelling of chloride transport in non-saturated concrete : from microscale to macroscale

Corrosion of steel bars embedded in concrete has a great influence on structural performance and durability of reinforced concrete. Chloride penetration is considered to be a primary cause of concrete deterioration in a vast majority of structures. Therefore, modelling of chloride penetration into concrete has become an area of great interest. The present work focuses on modelling of chloride transport in concrete. The differential macroscopic equations which govern the problem were derived from the equations at the microscopic scale by comparing the porous network with a single equivalent pore whose properties are the same as the average properties of the real porous network. The resulting transport model, which accounts for diffusion, migration, advection, chloride binding and chloride precipitation, consists of three coupled differential equations. The first equation models the transport of chloride ions, while the other two model the flow of the pore water and the heat transfer. In order to calibrate the model, the material parameters to determine experimentally were identified. The differential equations were solved by means of the finite element method. The classical Galerkin method was employed for the pore solution flow and the heat transfer equations, while the streamline upwind Petrov Galerkin method was adopted for the transport equation in order to avoid spatial instabilities for advection dominated problems. The finite element codes are implemented in Matlab® . To retrieve a good understanding of the influence of each variable and parameter, a detailed sensitivity analysis of the model was carried out. In order to determine the diffusive and hygroscopic properties of the studied concretes, as well as their chloride binding capacity, an experimental analysis was performed. The model was successfully compared with experimental data obtained from an offshore oil platform located in Brazil. Moreover, apart from the main objectives, numerous results were obtained throughout this work. For instance, several diffusion coefficients and the relation between them are discussed. It is shown how the electric field set up between the ionic species depends on the gradient of the species’ concentrations. Furthermore, the capillary hysteresis effects are illustrated by a proposed model, which leads to the determination of several microstructure properties, such as the pore size distribution and the tortuosity-connectivity of the porous network. El fenómeno de corrosión del acero de refuerzo embebido en el hormigón ha tenido gran influencia en estructuras de hormigón armado, tanto en su funcionalidad estructural como en aspectos de durabilidad. La penetración de cloruros en el interior del hormigón esta considerada como el factor principal en el deterioro de la gran mayoría de estructuras. Por lo tanto, la modelización numérica de dicho fenómeno ha generado gran interés. El presente trabajo de investigación se centra en la modelización del transporte de cloruros en el interior del hormigón. Las ecuaciones diferenciales que gobiernan los fenómenos a nivel macroscópico se deducen de ecuaciones planteadas a nivel microscópico. Esto se obtiene comparando la red porosa con un poro equivalente, el cual mantiene las mismas propiedades de la red porosa real. El modelo está constituido por tres ecuaciones diferenciales acopladas que consideran el transporte de cloruros, el flujo de la solución de poro y la transferencia de calor. Con estas ecuaciones se tienen en cuenta los fenómenos de difusión, migración, advección, combinación y precipitación de cloruros. El análisis llevado a cabo en este trabajo ha definido los parámetros necesarios para calibrar el modelo. De acuerdo con ellas, se seleccionaron los ensayos experimentales a realizar. Las ecuaciones diferenciales se resolvieron mediante el método de elementos finitos. El método clásico de Galerkin se empleó para solucionar las ecuaciones de flujo de la solución de poro y de la transferencia de calor, mientras que el método streamline upwind Petrov-Galerkin se utilizó para resolver la ecuación de transporte de cloruros con la finalidad de evitar inestabilidades espaciales en problemas con advección dominante. El código de elementos finitos está implementado en Matlab® . Con el objetivo de facilitar la comprensión del grado de influencia de cada variable y parámetro, se realizó un análisis de sensibilidad detallado del modelo. Se llevó a cabo una campaña experimental sobre los hormigones estudiados, con el objeto de obtener sus propiedades difusivas, químicas e higroscópicas. El modelo se contrastó con datos experimentales obtenidos en una plataforma petrolera localizada en Brasil. Las simulaciones numéricas corroboraron los datos experimentales. Además, durante el desarrollo de la investigación se obtuvieron resultados paralelos a los planteados inicialmente. Por ejemplo, el análisis de diferentes coeficientes de difusión y la relación entre ellos. Así como también se observó que el campo eléctrico establecido entre las especies iónicas disueltas en la solución de poro depende del gradiente de concentración de las mismas. Los efectos de histéresis capilar son expresados por el modelo propuesto, el cual conduce a la determinación de una serie de propiedades microscópicas, tales como la distribución del tamaño de poro, además de la tortuosidad y conectividad de la red porosa.

Development of a methodology and tool to evaluate the impact of ICT measures on road transport emissions

The paper presents the main elements of a project entitled ICT-Emissions that aims at developing a novel methodology to evaluate the impact of ICT-related measures on mobility, vehicle energy consumption and CO2 emissions of vehicle fleets at the local scale, in order to promote the wider application of the most appropriate ICT measures. The proposed methodology combines traffic and emission modelling at micro and macro scales. These will be linked with interfaces and submodules which will be specifically designed and developed. A number of sources are available to the consortium to obtain the necessary input data. Also, experimental campaigns are offered to fill in gaps of information in traffic and emission patterns. The application of the methodology will be demonstrated using commercially available software. However, the methodology is developed in such a way as to enable its implementation by a variety of emission and traffic models. Particular emphasis is given to (a) the correct estimation of driver behaviour, as a result of traffic-related ICT measures, (b) the coverage of a large number of current vehicle technologies, including ICT systems, and (c) near future technologies such as hybrid, plug-in hybrids, and electric vehicles. The innovative combination of traffic, driver, and emission models produces a versatile toolbox that can simulate the impact on energy and CO2 of infrastructure measures (traffic management, dynamic traffic signs, etc.), driver assistance systems and ecosolutions (speed/cruise control, start/stop systems, etc.) or a combination of measures (cooperative systems).The methodology is validated by application in the Turin area and its capacity is further demonstrated by application in real world conditions in Madrid and Rome.

Effect of solute and matric potential on growth rate of fungal species isolated from cheese

The effect of water potential ( J w ) on the growth of 15 fungal species isolated from cheeses was analysed. The species, identified mainly by analysis of DNA sequences, belonged to genera Penicillium , Geotrichum , Mucor , Aspergillus , Microascus and Talaromyces . Particularly, the effect of matric potential ( J m ), and ionic (NaCl) and non-ionic (glycerol) solute potentials ( J s ) on growth rate was studied. The response of strains was highly dependent on the type of J w . For J s , clear profiles for optimal, permissive and marginal conditions for growth were obtained, and differences in growth rate were achieved comparing NaCl and glycerol for most of the species. Conversely, a sustained growth was obtained for J m in all the strains, with the exception of Aspergillus pseudoglaucus , whose growth increased proportionally to the level of water stress. Our results might help to understand the impact of environmental factors on the ecophysiology and dynamics of fungal populations associated to cheeses.

Calibration of WAVE in irrigated maize: fallow vs. cover crops.

Nitrate leaching decreases crop available N and increases water contamination. Replacing fallow by cover crops (CC) is an alternative to reduce nitrate contamination, because it reduces overall drainage and soil mineral N accumulation. A study of the soil N and nitrate leaching was conducted during 5 years in a semi-arid irrigated agricultural area of Central Spain. Three treatments were studied during the intercropping period of maize (Zea mays L.): barley (Hordeum vulgare L.), vetch (Vicia villosa L.), and fallow. Cover crops, sown in October, were killed by glyphosate application in March, allowing direct seeding of maize in April. All treatments were irrigated and fertilised following the same procedure. Soil water content was measured using capacity probes. Soil Nmin accumulation was determined along the soil profile before sowing and after harvesting maize. Soil analysis was conducted at six depths every 0.20m in each plot in samples from 0 to 1.2-m depth. The mechanistic water balance model WAVE was applied in order to calculate drainage and plant growth of the different treatments, and apply them to the N balance. We evaluated the water balance of this model using the daily soil water content measurements of this field trial. A new Matlab version of the model was evaluated as well. In this new version improvements were made in the solute transport module and crop module. In addition, this new version is more compatible with external modules for data processing, inverse calibration and uncertainty analysis than the previous Fortran version. The model showed that drainage during the irrigated period was minimized in all treatments, because irrigation water was adjusted to crop needs, leading to nitrate accumulation on the upper layers after maize harvest. Then, during the intercrop period, most of the nitrate leaching occurred. Cover crops usually led to a shorter drainage period, lower drainage water amount and lower nitrate leaching than the treatment with fallow. These effects resulted in larger nitrate accumulation in the upper layers of the soil after CC treatments.

Stability and mechanical evaluation of bovine pericardium cross-linked with polyurethane prepolymer in aqueous medium

The present study investigates the potential use of non-catalyzed water-soluble blocked polyurethane prepolymer (PUP) as a bifunctional cross-linker for collagenous scaffolds. The effect of concentration (5, 10, 15 and 20%), time (4, 6, 12 and 24 h), medium volume (50, 100, 200 and 300%) and pH (7.4, 8.2, 9 and 10) over stability, microstructure and tensile mechanical behavior of acellular pericardial matrix was studied. The cross-linking index increased up to 81% while the denaturation temperature increased up to 12 °C after PUP crosslinking. PUP-treated scaffold resisted the collagenase degradation (0.167 ± 0.14 mmol/g of liberated amine groups vs. 598 ± 60 mmol/g for non-cross-linked matrix). The collagen fiber network was coated with PUP while viscoelastic properties were altered after cross-linking. The treatment of the pericardial scaffold with PUP allows (i) different densities of cross-linking depending of the process parameters and (ii) tensile properties similar to glutaraldehyde method.

Road fright transport decoupling: A comparative analysis between the United Kingdom and Spain

Economic growth has traditionally been linked to road freight transport demand, leading to a steady rise in social and environmental impacts. Concern about this problem has caused the EU to promote a decoupling strategy aimed at boosting sustainable development in European countries by improving the efficiency of transport systems without curbing economic growth. Over the last few years empirical evidence in some countries such as the United Kingdom has shown an increase in GDP while the volume of road freight traffic has remained stable or even decreased. This paper compares recent decoupling trends by analyzing the evolution of road tonne-kms/GDP relationship in the United Kingdom and Spain from 1999 to 2007. This comparison seeks to identify the main differences and key drivers of decoupling in both countries. We first provide an overview of the divergences between both economic structures and levels of road transport intensity. Then we conduct a decomposition analysis in order to identify the variables that explain the evolution of truck traffic per unit of GDP in each country. The results show that the increasing share of services in GDP has substantially contributed to decreasing road transport demand in both cases. Changes in road transport intensity due to improvements in logistic and supply chain management have been more successful in the UK than in Spain.

The impact of the economic crisis and policy actions on GHG emissions from road transport in Spain

Road traffic is the greatest contributor to the carbon footprint of the transport sector and reducing it has become one of the main targets of sustainable transport policies. An analysis of the main factors influencing greenhouse gas (GHG) emissions is essential for designing new energy- and environmentally efficient strategies for the road transport. This paper addresses this need by (i) identifying factors which influence the carbon footprint, including traffic activity, fuel economy and socioeconomic development; and (ii) proposing a methodological framework which uses Modified Laspeyres Index decomposition to analyze the effect of important drivers on the changes in emissions of road transport in Spain during the period from 1990 to 2010. The results demonstrate that the country׳s economic growth has been closely linked to the rise in GHG emissions. The innovative contribution of this paper is the special analysis of the changes in mobility patterns and GHG emissions during the economic crisis, when, for the first time, Spanish road traffic emissions decreased. The reduction of road transport and improved energy efficiency has been powerful contributors to this decrease, demonstrating the effectiveness of energy-saving measures. On the basis of this analysis, several tailored policy recommendations have been suggested for future implementation.

Implications of agricultural production, policy and land use changes on water resource assessment

Una gestión más eficiente y equitativa del agua a escala de cuenca no se puede centrar exclusivamente en el recurso hídrico en sí, sino también en otras políticas y disciplinas científicas. Existe un consenso creciente de que, además de la consideración de las cambiantes condiciones climáticas, es necesaria una integración de ámbitos de investigación tales como la agronomía, planificación del territorio y ciencias políticas y económicas a fin de satisfacer de manera sostenible las demandas de agua por parte de la sociedad y del medio natural. La Política Agrícola Común (PAC) es el principal motor de cambio en las tendencias de paisajes rurales y sistemas agrícolas, pero el deterioro del medio ambiente es ahora una de las principales preocupaciones. Uno de los cambios más relevantes se ha producido con la expansión e intensificación del olivar en España, principalmente con nuevas zonas de regadío o la conversión de olivares de secano a sistemas en regadío. Por otra parte, el cambio de las condiciones climáticas podría ejercer un papel importante en las tendencias negativas de las aportaciones a los ríos, pero no queda claro el papel que podrían estar jugando los cambios de uso de suelo y cobertura vegetal sobre las tendencias negativas de caudal observadas. Esta tesis tiene como objetivo mejorar el conocimiento de los efectos de la producción agrícola, política agraria y cambios de uso de suelo y cobertura vegetal sobre las condiciones de calidad del agua, respuesta hidrológica y apropiación del agua por parte de la sociedad. En primer lugar, el estudio determina las tendencias existentes de nitratos y sólidos en suspensión en las aguas superficiales de la cuenca del río Guadalquivir durante el periodo de 1998 a 2009. Desde una perspectiva de política agraria, la investigación trata de evaluar mediante un análisis de datos de panel las principales variables, incluyendo la reforma de la PAC de 2003, que están teniendo una influencia en ambos indicadores de calidad. En segundo lugar, la apropiación del agua y el nivel de contaminación por nitratos debido a la producción del aceite de oliva en España se determinan con una evaluación de la huella hídrica (HH), teniendo en cuenta una variabilidad espacial y temporal a largo de las provincias españolas y entre 1997 y 2008. Por último, la tesis analiza los efectos de los cambios de uso de suelo y cobertura vegetal sobre las tendencias negativas observadas en la zona alta del Turia, cabecera de la cuenca del río Júcar, durante el periodo 1973-2008 mediante una modelización ecohidrológica. En la cuenca del Guadalquivir cerca del 20% de las estaciones de monitoreo muestran tendencias significativas, lineales o cuadráticas, para cada indicador de calidad de agua. La mayoría de las tendencias significativas en nitratos están aumentando, y la mayoría de tendencias cuadráticas muestran un patrón en forma de U. Los modelos de regresión de datos de panel muestran que las variables más importantes que empeoran ambos indicadores de calidad del agua son la intensificación de biomasa y las exportaciones de ambos indicadores de calidad procedentes de aguas arriba. En regiones en las que el abandono agrícola y/o desintensificación han tenido lugar han mejorado las condiciones de calidad del agua. Para los nitratos, el desacoplamiento de las subvenciones a la agricultura y la reducción de la cuantía de las subvenciones a tierras de regadío subyacen en la reducción observada de la concentración de nitratos. Las medidas de modernización de regadíos y el establecimiento de zonas vulnerables a nitratos reducen la concentración en subcuencas que muestran una tendencia creciente de nitratos. Sin embargo, el efecto de las exportaciones de nitratos procedente de aguas arriba, la intensificación de la biomasa y los precios de los cultivos presentan un mayor peso, explicando la tendencia creciente observada de nitratos. Para los sólidos en suspensión, no queda de forma evidente si el proceso de desacoplamiento ha influido negativa o positivamente. Sin embargo, los mayores valores de las ayudas agrarias aún ligadas a la producción, en particular en zonas de regadío, conllevan un aumento de las tasas de erosión. Aunque la cuenca del Guadalquivir ha aumentado la producción agrícola y la eficiencia del uso del agua, el problema de las altas tasas de erosión aún no ha sido mitigado adecuadamente. El estudio de la huella hídrica (HH) revela que en 1 L de aceite de oliva español más del 99,5% de la HH está relacionado con la producción de la aceituna, mientras que menos del 0,5% se debe a otros componentes, es decir, a la botella, tapón y etiqueta. Durante el período estudiado, la HH verde en secano y en regadío representa alrededor del 72% y 12%, respectivamente, del total de la HH. Las HHs azul y gris representan 6% y 10%, respectivamente. La producción de aceitunas se concentra en regiones con una HH menor por unidad de producto. La producción de aceite de oliva ha aumentado su productividad del agua durante 1997-2008, incentivado por los crecientes precios del aceite, como también lo ha hecho la cantidad de exportaciones de agua virtual. De hecho, las mayores zonas productoras presentan una eficiencia alta del uso y de productividad del agua, así como un menor potencial de contaminación por nitratos. Pero en estas zonas se ve a la vez reflejado un aumento de presión sobre los recursos hídricos locales. El aumento de extracciones de agua subterránea relacionadas con las exportaciones de aceite de oliva podría añadir una mayor presión a la ya estresada cuenca del Guadalquivir, mostrando la necesidad de equilibrar las fuerzas del mercado con los recursos locales disponibles. Los cambios de uso de suelo y cobertura vegetal juegan un papel importante en el balance del agua de la cuenca alta del Turia, pero no son el principal motor que sustenta la reducción observada de caudal. El aumento de la temperatura es el principal factor que explica las mayores tasas de evapotranspiración y la reducción de caudales. Sin embargo, los cambios de uso de suelo y el cambio climático han tenido un efecto compensatorio en la respuesta hidrológica. Por un lado, el caudal se ha visto afectado negativamente por el aumento de la temperatura, mientras que los cambios de uso de suelo y cobertura vegetal han compensado positivamente con una reducción de las tasas de evapotranspiración, gracias a los procesos de disminución de la densidad de matorral y de degradación forestal. El estudio proporciona una visión que fortalece la interdisciplinariedad entre la planificación hidrológica y territorial, destacando la necesidad de incluir las implicaciones de los cambios de uso de suelo y cobertura vegetal en futuros planes hidrológicos. Estos hallazgos son valiosos para la gestión de la cuenca del río Turia, y el enfoque empleado es útil para la determinación del peso de los cambios de uso de suelo y cobertura vegetal en la respuesta hidrológica en otras regiones. ABSTRACT Achieving a more efficient and equitable water management at catchment scale does not only rely on the water resource itself, but also on other policies and scientific knowledge. There is a growing consensus that, in addition to consideration of changing climate conditions, integration with research areas such as agronomy, land use planning and economics and political science is required to meet sustainably the societal and environmental water demands. The Common Agricultural Policy (CAP) is a main driver for trends in rural landscapes and agricultural systems, but environmental deterioration is now a principal concern. One of the most relevant changes has occurred with the expansion and intensification of olive orchards in Spain, taking place mainly with new irrigated areas or with the conversion from rainfed to irrigated systems. Moreover, changing climate conditions might exert a major role on water yield trends, but it remains unclear the role that ongoing land use and land cover changes (LULCC) might have on observed river flow trends. This thesis aims to improve the understanding of the effects of agricultural production, policies and LULCC on water quality conditions, hydrological response and human water appropriation. Firstly, the study determines the existing trends for nitrates and suspended solids in the Guadalquivir river basin’s surface waters (south Spain) during the period from 1998 to 2009. From a policy perspective, the research tries to assess with panel data analysis the main drivers, including the 2003 CAP reform, which are having an influence on both water quality indicators. Secondly, water appropriation and nitrate pollution level originating from the production of olive oil in Spain is determined with a water footprint (WF) assessment, considering a spatial temporal variability across the Spanish provinces and from 1997 to 2008 years. Finally, the thesis analyzes the effects of the LULCC on the observed negative trends over the period 1973-2008 in the Upper Turia basin, headwaters of the Júcar river demarcation (east Spain), with ecohydrological modeling. In the Guadalquivir river basin about 20% of monitoring stations show significant trends, linear or quadratic, for each water quality indicator. Most significant trends of nitrates are augmenting than decreasing, and most significant quadratic terms of both indicators exhibit U-shaped patterns. The panel data models show that the most important drivers that are worsening nitrates and suspended solids in the basin are biomass intensification and exports of both water quality indicators from upland regions. In regions that agricultural abandonment and/or de-intensification have taken place the water quality conditions have improved. For nitrates, the decoupling of agricultural subsidies and the reduction of the amount of subsidies to irrigated land underlie the observed reduction of nitrates concentration. Measures of irrigation modernization and establishment of vulnerable zones to nitrates ameliorate the concentration of nitrates in subbasins showing an increasing trend. However, the effect of nitrates load from upland areas, intensification of biomass and crop prices present a greater weight leading to the final increasing trend in this subbasins group, where annual crops dominate. For suspended solids, there is no clear evidence that decoupling process have influenced negatively or positively. Nevertheless, greater values of subsidies still linked to production, particularly in irrigated regions, lead to increasing erosion rates. Although agricultural production has augmented in the basin and water efficiency in the agricultural sector has improved, the issue of high erosion rates has not yet been properly faced. The water footprint (WF) assessment reveals that for 1 L Spanish olive oil more than 99.5% of the WF is related to the olive fruit production, whereas less than 0.5% is due to other components i.e. bottle, cap and label. Over the studied period, the green WF in rainfed and irrigated systems represents about 72% and 12%, respectively, of the total WF. Blue and grey WFs represent 6% and 10%, respectively. The olive production is concentrated in regions with the smallest WF per unit of product. The olive oil production has increased its apparent water productivity from 1997 to 2008 incentivized by growing trade prices, but also did the amount of virtual water exports. In fact, the largest producing areas present high water use efficiency per product and apparent water productivity as well as less nitrates pollution potential, but this enhances the pressure on the available water resources. Increasing groundwater abstractions related to olive oil exports may add further pressure to the already stressed Guadalquivir basin. This shows the need to balance the market forces with the available local resources. Concerning the effects of LULCC on the Upper Turia basin’s streamflow, LULCC play a significant role on the water balance, but it is not the main driver underpinning the observed reduction on Turia's streamflow. Increasing mean temperature is the main factor supporting larger evapotranspiration rates and streamflow reduction. In fact, LULCC and climate change have had an offsetting effect on the streamflow generation during the study period. While streamflow has been negatively affected by increasing temperature, ongoing LULCC have positively compensated with reduced evapotranspiration rates, thanks to mainly shrubland clearing and forest degradation processes. The research provides insight for strengthening the interdisciplinarity between hydrological and spatial planning, highlighting the need to include the implications of LULCC in future hydrological plans. These findings are valuable for the management of the Turia river basin, as well as a useful approach for the determination of the weight of LULCC on the hydrological response in other regions.

Effect of solute and matric potential on growth rate of fungal species isolated from cheese

The effect of water potential ( J w ) on the growth of 15 fungal species isolated from cheeses was analysed. The species, identi fi ed mainly by analysis of DNA sequences, belonged to genera Penicillium, Geotrichum, Mucor , Aspergillus , Microascus and Talaromyces . Particularly, the effect of matric potential ( J m ), and ionic (NaCl) and non-ionic (glycerol) solute potentials ( J s ) on growth rate was studied. The response of strains was highly dependent on the type of J w . For J s, clear profiles for optimal, permissive and marginal conditions for growth were obtained, and differences in growth rate were achieved comparing NaCl and glycerol for most of the species. Conversely, a sustained growth was obtained for J m in all the strains, with the exception of Aspergillus pseudoglaucus, whose growth increased proportionally to the level of water stress. Our results might help to understand the impact of environmental factors on the ecophysiology and dynamics of fungal populations associated to cheeses.

Building resilience to water scarcity in Southern Spain: A case study of rice farming in Doñana protected wetlands

Agricultural water management needs to evolve in view of increased water scarcity, especially when farming and natural protected areas are closely linked. In the study site of Don?ana (southern Spain), water is shared by rice producers and a world heritage biodiversity ecosystem. Our aim is to contribute to defining adaptation strategies that may build resilience to increasing water scarcity and minimize water conflicts among agricultural and natural systems. The analytical framework links a participatory process with quantitative methods to prioritize the adaptation options. Bottom-up proposed adaptation measures are evaluated by a multi-criteria analysis (MCA) that includes both socioeconomic criteria and criteria of the ecosystem services affected by the adaptation options. Criteria weights are estimated by three different methods?analytic hierarchy process, Likert scale and equal weights?that are then compared. Finally, scores from an MCA are input into an optimization model used to determine the optimal land-use distribution in order to maximize utility and land-use diversification according to different scenarios of funds and water availability. While our results show a spectrum of perceptions of priorities among stakeholders, there is one overriding theme that is to define a way to restore part of the rice fields to natural wetlands. These results hold true under the current climate scenario and evenmore so under an increased water scarcity scenario.