Validation of microsatellite markers for cytotype discrimination in the model grass Brachypodium distachyon

Brachypodium distachyon (2n = 2x = 10) is a small annual grass species where the existence of three different cytotypes (10, 20 and 30 chromosomes) has long been regarded as a case of autopolyploid series, with x = 5. However, it has been demonstrated that the cytotypes assumed to be polyploids represent two separate Brachypodium species recently named as B. stacei (2n = 2x = 20) and B. hybridum (2n = 4x = 30). The aim of this study was to find a PCR-based alternative approach that could replace standard cytotyping methods (i. e., chromosome counting and flow cytometry) to characterize each of the three Brachypodium species. We have analyzed with four microsatellite (SSR) markers eighty-three Brachypodium distachyon-type lines from varied locations in Spain, including the Balearic and Canary Islands. Within this set of lines, 64, 4 and 15 had 10, 20 and 30 chromosomes, respectively. The surveyed markers produced cytotype-specific SSR profiles. So, a single amplification product was generated in the diploid samples, with non-overlapping allelic ranges between the 2n = 10 and 2n = 20 cytotypes, whereas two bands, one in the size range of each of the diploid cytotypes, were amplified in the 2n = 30 lines. Furthermore, the remarkable size difference obtained with the SSR ALB165 allowed the identification of the Brachypodium species by simple agarose gel electrophoresis.

Rapid sizing tool for a low cost university microsatellite structure subsystem

In recent decays university class small satellites are creating many opportunities for space research and professional trainings while at the same time responding to constrained budgets. In this work the main focus is on developing a simple and rapid structural sizing tool considering the main objectives of a low cost university class microsatellite project. In satellite projects, structure subsystem is one of the influential subsystems as a driver of the cost and acceptance of the final design. At the first steps of such projects there is no confirmed data regarding the launch vehicle or even in some cases there is no data for the satellite payload. Due to these facts, developing simple sizing tools at conceptual design phase for obtaining an over view of the effect of different variables is useful before entering complex calculations in detailed design phases. In this study, after developing a simple analytical model of satellite structure subsystem, a design space is evaluated with practical boundaries considering mass and dimensions constraints of such projects. The results are useful to give initial insight to establish the system level structural sizing

Clones Identification and Genetic Characterization of Garnacha Grapevine by Means of Different PCR-Derived Marker Systems

This study uses PCR-derived marker systems to investigate the extent and distribution of genetic variability of 53 Garnacha accessions coming from Italy, France and Spain. The samples studied include 28 Italian accessions (named Tocai rosso in Vicenza area; Alicante in Sicily and Elba island; Gamay perugino in Perugia province; Cannonau in Sardinia), 19 Spanish accessions of different types (named Garnacha tinta, Garnacha blanca, Garnacha peluda, Garnacha roja, Garnacha erguida, Garnacha roya) and 6 French accessions (named Grenache and Grenache noir). In order to verify the varietal identity of the samples, analyses based on 14 simple sequence repeat (SSR) loci were performed. The presence of an additional allele at ISV3 locus (151 bp) was found in four Tocai rosso accessions and in a Sardinian Cannonau clone, that are, incidentally, chimeras. In addition to microsatellite analysis, intravarietal variability study was performed using AFLP, SAMPL and M-AFLP molecular markers. AFLPs could discriminate among several Garnacha samples; SAMPLs allowed distinguishing few genotypes on the basis of their geographic origin, whereas M-AFLPs revealed plant-specific markers, differentiating all accessions. Italian samples showed the greatest variability among themselves, especially on the basis of their different provenance, while Spanish samples were the most similar, in spite of their morphological diversity.

Uso de marcadores moleculares para la racionalizaci��n y evaluaci��n de la calidad en las colecciones de Recursos Fitogen��ticos de trigo

Esta tesis tiene dos objetivos generales: el primero, analizar el uso de prote��nas del endospermo y SSRs para la racionalizaci��n de las colecciones de trigo, y el segundo, estudiar la influencia de las prote��nas del endospermo, del a��o de cultivo y del abonado nitrogenado en la calidad en un grupo de variedades locales espa��olas. Dentro del primer objetivo, se estudi�� la diversidad gen��tica de la colecci��n de Triticum monococcum L. (esca��a menor), y de una muestra de la colecci��n de Triticum turgidum L. (trigo duro) del CRF-INIA, con 2 y 6 loci de gliadinas, y 6 y 24 SSRs, para la esca��a menor y el trigo duro, respectivamente. Ambas colecciones presentaron una gran diversidad gen��tica, con una gran diferenciaci��n entre las variedades y peque��a dentro de ellas. Los loci de gliadinas mostraron una gran variabilidad, siendo los loci Gli-2 los m��s ��tiles para distinguir variedades. En la esca��a menor, las gliadinas presentaron mayor poder de discriminaci��n que los SSRs; aunque en trigo duro los SSRs identificaron m��s genotipos. El n��mero de alelos encontrado fue alto; 24 y 38 en gliadinas, y 29 y 203 en SSRs, en esca��a menor y trigo duro, respectivamente. En trigo duro, se identificaron 17 alelos nuevos de gliadinas lo que demuestra que el germoplasma espa��ol es muy singular. En ambas especies, se detectaron asociaciones entre la variaci��n al��lica en prolaminas y el origen geogr��fico y filogen��tico de las variedades. La utilidad de las prote��nas (6 loci de gliadinas, 2 loci de gluteninas y prote��na total) y de los SSRs (24 loci) para verificar duplicados, y analizar la variabilidad intraaccesi��n, se estudi�� en 23 casos de duplicados potenciales de trigo duro. Los resultados indicaron que tanto los biotipos como las accesiones duplicadas mostraban el mismo genotipo en gliadinas, pocas diferencias o ninguna en las subunidades de gluteninas HMW y prote��na total, y diferencias en menos de tres loci de SSRs. El mismo resultado se obtuvo para los biotipos de la colecci��n de T. monococcum. Sin embargo, las discrepancias observadas en algunos casos entre prote��nas y SSRs demostraron la utilidad del uso conjunto de ambos tipos de marcadores. Tanto las prote��nas como los SSRs mostraron gran concordancia con los caracteres agro-morfol��gicos, especialmente cuando las diferencias entre los genotipos eran grandes. Sin embargo, los caracteres agro-morfol��gicos fueron menos discriminantes que los marcadores moleculares. Para el segundo objetivo de la tesis, se analiz�� la variaci��n al��lica en siete loci de prolaminas relacionados con la calidad en trigo duro: Glu-A1 y Glu-B1 de gluteninas HMW, Glu-A3, Glu-B3 y Glu-B2 de gluteninas B-LMW, y Gli-A1 y Gli-B1 de gliadinas. La submuestra analizada inclu��a variedades locales de todas las provincias espa��olas donde se ha cultivado tradicionalmente el trigo duro. Todos los loci, excepto el Glu-B2, mostraron gran variabilidad gen��tica, siendo los Glu-3 los m��s polim��rficos. En total, se identificaron 65 alelos, de los que 29 eran nuevos, que representan una fuente importante de variabilidad gen��tica para la mejora de la calidad. Se detectaron diferencias en la composici��n en prolaminas entre la convar. turgidum y la zona norte, y la convar. durum y la zona sur; el genotipo Glu-B3new-1 - Gli-B1new-1 fue muy com��n en la convar. turgidum, mientras que el Glu-B3a - Gli-B1c, asociado con mejor calidad, fue m��s frecuente en la convar. durum. En la convar. turgidum, se observ�� mayor variabilidad que en la convar. durum, principalmente en los loci Glu-B1 y Glu-B3, lo que indica que esta convariedad puede ser una fuente valiosa de nuevos alelos de gluteninas. Esta submuestra fue evaluada para calidad (contenido en prote��na, P, y test de sedimentaci��n, SDSS) con dos dosis de abonado nitrogenado (N), y en dos a��os diferentes. No se detectaron interacciones Variedad �� A��o, ni Variedad �� N en la calidad. Para la P, los efectos ambientales (a��o y N) fueron mayores que el efecto de la variedad, siendo, en general, mayor la P con dosis altas de N. La variedad influy�� m��s en el test SDSS, que no se vio afectado por el a��o ni el N. El aumento del contenido en prote��na no influy�� significativamente sobre la fuerza del gluten estimada con el SDSS. Respecto a la influencia de las prolaminas en la fuerza del gluten, se confirm�� la superioridad del Glu-B3a; aunque tambi��n se detect�� una influencia alta y positiva de los alelos nuevos Glu-A3new-1, y Glu-B3new-6 y new-9. La no correlaci��n entre el rendimiento (evaluado en un trabajo anterior) y la P, en las variedades adaptadas a bajo N, permiti�� seleccionar cuatro variedades locales con alto rendimiento y buena fuerza del gluten para producci��n con bajo N. SUMMARY There are two main objectives in this thesis: The first, to analyse the use of endosperm proteins and SSRs to rationalize the wheat collections, and the second, to study the influence on quality of endosperm proteins, year and nitrogen fertilization in a group of Spanish landraces. For the first objective, we studied the genetic diversity of the collection of Triticum monococcum L. (cultivated einkorn), and of a sample of the collection of Triticum turgidum L. (durum wheat) maintained at the CRF-INIA. Two and 6 gliadin loci, and 6 and 24 SSRs, were used for einkorn and durum wheat, respectively. Both collections possessed a high genetic diversity, being the differentiation large between varieties and small within them. Gliadin loci showed great variability, being the loci Gli-2 the most useful for distinguish among varieties. In einkorn, the gliadins showed higher discrimination power than SSRs; although SSRs identified more genotypes in durum wheat. Large number of alleles were found; 24 and 38 in gliadins, and 29 and 203 in SSRs, for einkorn and durum wheat, respectively. In durum wheat, 17 new alleles of gliadins were identified, which indicate that Spanish durum wheat germplasm is rather unique. Some associations between prolamin alleles and geographical and phylogenetic origin of varieties were found in both species. The value of endosperm proteins (6 gliadin loci, 2 glutenin loci and total protein) and SSRs (24 loci) for validation of duplicates, and monitoring the intra-accession variability, was studied in 23 potential duplicates of durum wheat. The results indicated that biotypes and duplicated accessions showed identical gliadin genotype, few or none differences in HMW glutenin subunits and total protein, and less than three different SSR loci. A similar result was obtained for biotypes of T. monococcum. However, the discrepancies in some cases support the convenience to use together both marker systems. A good concordance among endosperm proteins, agro-morphological traits and SSRs were also found, mainly when differences between genotypes were high. However, agro-morphological traits discriminated less between accessions than molecular markers. For the second objective of the thesis, we analysed the allelic variation at seven prolamin loci, involved in durum wheat quality: Glu-A1 and Glu-B1 of HMW glutenin, Glu-A3, Glu-B3 and Glu-B2 of B-LMW glutenin, and Gli-A1 and Gli-B1 of gliadin. The subsample analysed included landraces from all the Spanish provinces where the crop was traditionally cultivated. All the loci, except for Glu-B2, showed high genetic variability, being Glu-3 the most polymorphic. A total of 65 alleles were studied, 29 of them being new, which represent an important source of variability for quality improvement. Differences in prolamin composition were detected between convar. turgidum and the North zone, and the convar. durum and the South zone; the genotype Glu-B3new-1 - Gli-B1new-1 was very common in the convar. turgidum, while the Glu- B3a - Gli-B1c, associated with better quality, was more frequent in the convar. durum. Higher variability was detected in the convar. turgidum than in the convar. durum, mainly at the Glu-B1 and Glu-B3, showing that this convariety could be a valuable source of new glutenin alleles. The subsample was evaluated for quality (protein content, P, and sedimentation test, SDSS) with two doses of nitrogen fertiliser (N), and in two different years. No significant Variety x Year or Variety x Nitrogen interactions were detected. For P, environmental (year and N) effects were higher than variety effect, being P values , in general, larger with high dose of N. The variety exhibited a strong influence on SDSS test, which was not affected by year and N. Increasing values of P did not significantly influence on gluten strength, estimated with the SDSS. Respect to the prolamin effects on gluten strength, the superiority of Glu-B3a was confirmed; although a high positive effect of the new alleles Glu-A3new-1, and Glu-B3new-6 and new-9 was also detected. The no correlation between yield (evaluated in a previous research) and P, in the landraces adapted to low N, allowed to select four landraces with high yield and high gluten strength for low N production.

Multiobjective optimization applied to structural sizing of low cost university-class microsatellite projects

In recent years, there has been continuing interest in the participation of university research groups in space technology studies by means of their own microsatellites. The involvement in such projects has some inherent challenges, such as limited budget and facilities. Also, due to the fact that the main objective of these projects is for educational purposes, usually there are uncertainties regarding their in orbit mission and scientific payloads at the early phases of the project. On the other hand, there are predetermined limitations for their mass and volume budgets owing to the fact that most of them are launched as an auxiliary payload in which the launch cost is reduced considerably. The satellite structure subsystem is the one which is most affected by the launcher constraints. This can affect different aspects, including dimensions, strength and frequency requirements. In this paper, the main focus is on developing a structural design sizing tool containing not only the primary structures properties as variables but also the system level variables such as payload mass budget and satellite total mass and dimensions. This approach enables the design team to obtain better insight into the design in an extended design envelope. The structural design sizing tool is based on analytical structural design formulas and appropriate assumptions including both static and dynamic models of the satellite. Finally, a Genetic Algorithm (GA) multiobjective optimization is applied to the design space. The result is a Pareto-optimal based on two objectives, minimum satellite total mass and maximum payload mass budget, which gives a useful insight to the design team at the early phases of the design.

Interactions between water and triacylglycerols may explain faster aging rates in stored germplasm at low temperatures

The purpose of this research is to explore the extent and significance of possible interacting factors on the viability of stored germplasm. Our work begins with characterizing the kinetics of TAG and water phase changes in peanut (Arachis hypogaea) and Papaya (Carica papaya) seeds equilibrated to different water contents and stored at temperatures between -5 and -80��C. Water and TAG phase was measured using a Perkin Elmer Differential Scanning Calorimeter. Cytoplasm ultra-structure was visualized without chemical fixatives using low temperature scanning electron microscopy (cryo-SEM) performed with a Zeiss DSN 960 scanning microscope equipped with a Cryotrans CT-1500 cold plate (Oxford, UK).

Estudios termodin��micos de los procesos de vitrificaci��n en la crioconservaci��n de germoplasma vegetal=Thermodynamic studies of vitrification processes in plant germplasm cryopreservation

En la actualidad, las t��cnicas de crioconservaci��n poseen una importancia creciente para el almacenamiento a largo plazo de germoplasma vegetal. En las dos ��ltimas d��cadas, estos m��todos experimentaron un gran desarrollo y se han elaborado protocolos adecuados a diferentes sistemas vegetales, utilizando diversas estrategias como la vitrificaci��n, la encapsulaci��n-desecaci��n con cuentas de alginato y el m��todo de ���droplet���-vitrificaci��n. La presente tesis doctoral tiene como objetivo aumentar el conocimiento sobre los procesos implicados en los distintos pasos de un protocolo de crioconservaci��n, en relaci��n con el estado del agua presente en los tejidos y sus cambios, abordado mediante diversas t��cnicas biof��sicas, principalmente calorimetr��a diferencial de barrido (DSC) y microscop��a electr��nica de barrido a baja temperatura (crio-SEM). En un primer estudio sobre estos m��todos de crioconservaci��n, se describen las fases de enfriamiento hasta la temperatura del nitr��geno l��quido y de calentamiento hasta temperatura ambiente, al final del periodo de almacenamiento, que son cr��ticas para la supervivencia del material crioconservado. Tanto enfriamiento como calentamiento deben ser realizados lo m��s r��pidamente posible pues, aunque los bajos contenidos en agua logrados en etapas previas de los protocolos reducen significativamente las probabilidades de formaci��n de hielo, ��stas no son del todo nulas. En ese contexto, se analiza tambi��n la influencia de las velocidades de enfriamiento y calentamiento de las soluciones de crioconservaci��n de plantas en sus par��metros termof��sicos referente a la vitrificaci��n, en relaci��n su composici��n y concentraci��n de compuestos. Estas soluciones son empleadas en la mayor parte de los protocolos actualmente utilizados para la crioconservaci��n de material vegetal. Adem��s, se estudia la influencia de otros factores que pueden determinar la estabilidad del material vitrificado, tales como en envejecimiento del vidrio. Se ha llevado a cabo una investigaci��n experimental en el empleo del crio-SEM como una herramienta para visualizar el estado v��treo de las c��lulas y tejidos sometidos a los procesos de crioconservaci��n. Se ha comparado con la m��s conocida t��cnica de calorimetr��a diferencial de barrido, obteni��ndose resultados muy concordantes y complementarios. Se explor�� tambi��n por estas t��cnicas el efecto sobre tejidos vegetales de la adaptaci��n a bajas temperaturas y de la deshidrataci��n inducida por los diferentes tratamientos utilizados en los protocolos. Este estudio permite observar la evoluci��n biof��sica de los sistemas en el proceso de crioconservaci��n. Por ��ltimo, se estudi�� la aplicaci��n de pel��culas de quitosano en las cuentas de alginato utilizadas en el protocolo de encapsulaci��n. No se observaron cambios significativos en su comportamiento frente a la deshidrataci��n, en sus par��metros calorim��tricos y en la superficie de las cuentas. Su aplicaci��n puede conferir propiedades adicionales prometedoras. ABSTRACT Currently, cryopreservation techniques have a growing importance for long term plant germplasm storage. These methods have undergone great progress during the last two decades, and adequate protocols for different plant systems have been developed, making use of diverse strategies, such as vitrification, encapsulation-dehydration with alginate beads and the dropletvitrification method. This PhD thesis has the goal of increasing the knowledge on the processes underlying the different steps of cryopreservation protocols, in relation with the state of water on tissues and its changes, approached through diverse biophysical techniques, especially differential scanning calorimetry (DSC) and low-temperature scanning electron microscopy (cryo-SEM). The processes of cooling to liquid nitrogen temperature and warming to room temperature, at the end of the storage period, critical for the survival of the cryopreserved material, are described in a first study on these cryopreservation methods. Both cooling and warming must be carried out as quickly as possible because, although the low water content achieved during previous protocol steps significantly reduces ice formation probability, it does not completely disappear. Within this context, the influence of plant vitrification solutions cooling and warming rate on their vitrification related thermophysical parameters is also analyzed, in relation to its composition and component concentration. These solutions are used in most of the currently employed plant material cryopreservation protocols. Additionally, the influence of other factors determining the stability of vitrified material is studied, such as glass aging. An experimental research work has been carried out on the use of cryo-SEM as a tool for visualizing the glassy state in cells and tissues, submitted to cryopreservation processes. It has been compared with the better known differential scanning calorimetry technique, and results in good agreement and complementary have been obtained. The effect on plant tissues of adaptation to low temperature and of the dehydration induced by the different treatments used in the protocols was explored also by these techniques. This study allows observation of the system biophysical evolution in the cryopreservation process. Lastly, the potential use of an additional chitosan film over the alginate beads used in encapsulation protocols was examined. No significant changes could be observed in its dehydration and calorimetric behavior, as well as in its surface aspect; its application for conferring additional properties to gel beads is promising.

Creation and Validation of the Spanish Durum Wheat Core Collection.

Spanish wheat (Triticum spp.) landraces have a considerable polymorphism, containing many unique alleles, relative to other collections. The existence of a core collection is a favored approach for breeders to efficiently explore novel variation and enhance the use of germplasm. In this study, the Spanish durum wheat (Triticum turgidum L.) core collection (CC) was created using a population structure���based method, grouping accessions by subspecies and allocating the number of genotypes among populations according to the diversity of simple sequence repeat (SSR) markers. The CC of 94 genotypes was established, which accounted for 17% of the accessions in the entire collection. An alternative core collection (CH), with the same number of genotypes per subspecies and maximizing the coverage of SSR alleles, was assembled with the Core Hunter software. The quality of both core collections was compared with a random core collection and evaluated using geographic, agromorphological, and molecular marker data not previously used in the selection of genotypes. Both core collections had a high genetic representativeness, which validated their sampling strategies. Geographic and agromorphological variation, phenotypic correlations, and gliadin alleles of the original collection were more accurately depicted by the CC. Diversity arrays technology (DArT) markers revealed that the CC included genotypes less similar than the CH. Although more SSR alleles were retained by the CH (94%) than by the CC (91%), the results showed that the CC was better than CH for breeding purposes.

Thermographic imaging: assessment of drought and heat tolerance in Spanish germplasm of Brachypodium distachyon.

The annual grass Brachypodium distachyon has been recently recognized as the model plant for functional genomics of temperate grasses, including cereals of economic relevance like wheat and barley. Sixty-two lines of B. distachyon were assessed for response to drought stress and heat tolerance. All these lines, except the reference genotype BD21, derive from specimens collected in 32 distinct locations of the Iberian Peninsula, covering a wide range of geo- climatic conditions. Sixteen lines of Brachypodium hybridum, an allotetraploid closely related to B. distachyon were used as reference of abiotic-stress well-adapted genotypes. Drought tolerance was assessed in a green-house trial. At the rosette-stage, no irrigation was applied to treated plants whereas their replicates at the control were maintained well watered during all the experiment. Thermographic images of treated and control plants were taken after 2 and 3 weeks of drought treatment, when stressed plants showed medium and extreme wilting symptoms. The mean leaf temperature of stressed (LTs) and control (LTc) plants was estimated based upon thermographic records from selected pixels (183 per image) that strictly correspond to leaf tissue. The response to drought was based on the analysis of two parameters: LTs and the thermal difference (TD) between stressed and control plants (LTs ��� LTc). The response to heat stress was based on LTc. Comparison of the mean values of these parameters showed that: 1) Genotypes better adapted to drought (B. hybridum lines) presented a higher LTs and TD than B. distachyon lines. 2) Under high temperature conditions, watered plants of B. hybridum lines maintained lower LTc than those of B. distachyon. Those results suggest that in these species adaptation to drought is linked to a more efficient stomata regulation: under water stress stomata are closed, increasing foliar temperature but also water use efficiency by reducing transpiration. With high temperature and water availability the results are less definite, but still seems that opening stomata allow plants to increase transpiration and therefore to diminish foliar temperature.

On the simulation of the UPMSAT-2 microsatellite power

Simulation of satellite subsystems behaviour is extramely important in the design at early stages. The subsystems are normally simulated in the both ways : isolated and as part of more complex simulation that takes into account imputs from other subsystems (concurrent design). In the present work, a simple concurrent simulation of the power subsystem of a microsatellite, UPMSat-2, is described. The aim of the work is to obtain the performance profile of the system (battery charging level, power consumption by the payloads, power supply from solar panels....). Different situations such as battery critical low or high level, effects of high current charging due to the low temperature of solar panels after eclipse,DoD margins..., were analysed, and different safety strategies studied using the developed tool (simulator) to fulfil the mission requirements. Also, failure cases were analysed in order to study the robustness of the system. The mentioned simulator has been programed taking into account the power consumption performances (average and maximum consumptions per orbit/day) of small part of the subsystem (SELEX GALILEO SPVS modular generators built with Azur Space solar cells, SAFT VES16 6P4S Li-ion battery, SSBV magnetometers, TECNOBIT and DATSI/UPM On Board Data Handling -OBDH-...). The developed tool is then intended to be a modular simulator, with the chance of use any other components implementing some standard data.

Multidisciplinary Design Optimization Application to Conceptual Design of University-class Microsatellite Projects

Esta tesis se ha realizado en el contexto del proyecto UPMSat-2, que es un microsat��lite dise��ado, construido y operado por el Instituto Universitario de Microgravedad "Ignacio Da Riva" (IDR / UPM) de la Universidad Polit��cnica de Madrid. Aplicaci��n de la metodolog��a Ingenier��a Concurrente (Concurrent Engineering: CE) en el marco de la aplicaci��n de dise��o multidisciplinar (Multidisciplinary Design Optimization: MDO) es uno de los principales objetivos del presente trabajo. En los ��ltimos a��os, ha habido un inter��s continuo en la participaci��n de los grupos de investigaci��n de las universidades en los estudios de la tecnolog��a espacial a trav��s de sus propios microsat��lites. La participaci��n en este tipo de proyectos tiene algunos desaf��os inherentes, tales como presupuestos y servicios limitados. Adem��s, debido al hecho de que el objetivo principal de estos proyectos es fundamentalmente educativo, por lo general hay incertidumbres en cuanto a su misi��n en ��rbita y cargas ��tiles en las primeras fases del proyecto. Por otro lado, existen limitaciones predeterminadas para sus presupuestos de masa, volumen y energ��a, debido al hecho de que la mayor��a de ellos est��n considerados como una carga ��til auxiliar para el lanzamiento. De este modo, el costo de lanzamiento se reduce considerablemente. En este contexto, el subsistema estructural del sat��lite es uno de los m��s afectados por las restricciones que impone el lanzador. Esto puede afectar a diferentes aspectos, incluyendo las dimensiones, la resistencia y los requisitos de frecuencia. En la primera parte de esta tesis, la atenci��n se centra en el desarrollo de una herramienta de dise��o del subsistema estructural que eval��a, no s��lo las propiedades de la estructura primaria como variables, sino tambi��n algunas variables de nivel de sistema del sat��lite, como la masa de la carga ��til y la masa y las dimensiones extremas de sat��lite. Este enfoque permite que el equipo de dise��o obtenga una mejor visi��n del dise��o en un espacio de dise��o extendido. La herramienta de dise��o estructural se basa en las f��rmulas y los supuestos apropiados, incluyendo los modelos est��ticos y din��micos del sat��lite. Un algoritmo gen��tico (Genetic Algorithm: GA) se aplica al espacio de dise��o para optimizaciones de objetivo ��nico y tambi��n multiobjetivo. El resultado de la optimizaci��n multiobjetivo es un Pareto-optimal basado en dos objetivo, la masa total de sat��lites m��nimo y el m��ximo presupuesto de masa de carga ��til. Por otro lado, la aplicaci��n de los microsat��lites en misiones espaciales es de inter��s por su menor coste y tiempo de desarrollo. La gran necesidad de las aplicaciones de teledetecci��n es un fuerte impulsor de su popularidad en este tipo de misiones espaciales. Las misiones de tele-observaci��n por sat��lite son esenciales para la investigaci��n de los recursos de la tierra y el medio ambiente. En estas misiones existen interrelaciones estrechas entre diferentes requisitos como la altitud orbital, tiempo de revisita, el ciclo de vida y la resoluci��n. Adem��s, todos estos requisitos puede afectar a toda las caracter��sticas de dise��o. Durante los ��ltimos a��os la aplicaci��n de CE en las misiones espaciales ha demostrado una gran ventaja para llegar al dise��o ��ptimo, teniendo en cuenta tanto el rendimiento y el costo del proyecto. Un ejemplo bien conocido de la aplicaci��n de CE es la CDF (Facilidad Dise��o Concurrente) de la ESA (Agencia Espacial Europea). Est�� claro que para los proyectos de microsat��lites universitarios tener o desarrollar una instalaci��n de este tipo parece estar m��s all�� de las capacidades del proyecto. Sin embargo, la pr��ctica de la CE a cualquier escala puede ser beneficiosa para los microsat��lites universitarios tambi��n. En la segunda parte de esta tesis, la atenci��n se centra en el desarrollo de una estructura de optimizaci��n de dise��o multidisciplinar (Multidisciplinary Design Optimization: MDO) aplicable a la fase de dise��o conceptual de microsat��lites de teledetecci��n. Este enfoque permite que el equipo de dise��o conozca la interacci��n entre las diferentes variables de dise��o. El esquema MDO presentado no s��lo incluye variables de nivel de sistema, tales como la masa total del sat��lite y la potencia total, sino tambi��n los requisitos de la misi��n como la resoluci��n y tiempo de revisita. El proceso de dise��o de microsat��lites se divide en tres disciplinas; a) dise��o de ��rbita, b) dise��o de carga ��til y c) dise��o de plataforma. En primer lugar, se calculan diferentes par��metros de misi��n para un rango pr��ctico de ��rbitas helio-s��ncronas (sun-synchronous orbits: SS-Os). Luego, seg��n los par��metros orbitales y los datos de un instrumento como referencia, se calcula la masa y la potencia de la carga ��til. El dise��o de la plataforma del sat��lite se estima a partir de los datos de la masa y potencia de los diferentes subsistemas utilizando relaciones emp��ricas de dise��o. El dise��o del subsistema de potencia se realiza teniendo en cuenta variables de dise��o m��s detalladas, como el escenario de la misi��n y diferentes tipos de c��lulas solares y bater��as. El escenario se selecciona, de modo de obtener una banda de cobertura sobre la superficie terrestre paralelo al Ecuador despu��s de cada intervalo de revisita. Con el objetivo de evaluar las interrelaciones entre las diferentes variables en el espacio de dise��o, todas las disciplinas de dise��o mencionados se combinan en un c��digo unificado. Por ��ltimo, una forma b��sica de MDO se ajusta a la herramienta de dise��o de sistema de sat��lite. La optimizaci��n del dise��o se realiza por medio de un GA con el ��nico objetivo de minimizar la masa total de microsat��lite. Seg��n los resultados obtenidos de la aplicaci��n del MDO, existen diferentes puntos de dise��os ��ptimos, pero con diferentes variables de misi��n. Este an��lisis demuestra la aplicabilidad de MDO para los estudios de ingenier��a de sistema en la fase de dise��o conceptual en este tipo de proyectos. La principal conclusi��n de esta tesis, es que el dise��o cl��sico de los sat��lites que por lo general comienza con la definici��n de la misi��n y la carga ��til no es necesariamente la mejor metodolog��a para todos los proyectos de sat��lites. Un microsat��lite universitario, es un ejemplo de este tipo de proyectos. Por eso, se han desarrollado un conjunto de herramientas de dise��o para encarar los estudios de la fase inicial de dise��o. Este conjunto de herramientas incluye diferentes disciplinas de dise��o centrados en el subsistema estructural y teniendo en cuenta una carga ��til desconocida a priori. Los resultados demuestran que la m��nima masa total del sat��lite y la m��xima masa disponible para una carga ��til desconocida a priori, son objetivos conflictivos. En este contexto para encontrar un Pareto-optimal se ha aplicado una optimizaci��n multiobjetivo. Seg��n los resultados se concluye que la selecci��n de la masa total por sat��lite en el rango de 40-60 kg puede considerarse como ��ptima para un proyecto de microsat��lites universitario con carga ��til desconocida a priori. Tambi��n la metodolog��a CE se ha aplicado al proceso de dise��o conceptual de microsat��lites de teledetecci��n. Los resultados de la aplicaci��n del CE proporcionan una clara comprensi��n de la interacci��n entre los requisitos de dise��o de sistemas de sat��lites, tales como la masa total del microsat��lite y la potencia y los requisitos de la misi��n como la resoluci��n y el tiempo de revisita. La aplicaci��n de MDO se hace con la minimizaci��n de la masa total de microsat��lite. Los resultados de la aplicaci��n de MDO aclaran la relaci��n clara entre los diferentes requisitos de dise��o del sistema y de misi��n, as�� como que permiten seleccionar las l��neas de base para el dise��o ��ptimo con el objetivo seleccionado en las primeras fase de dise��o. ABSTRACT This thesis is done in the context of UPMSat-2 project, which is a microsatellite under design and manufacturing at the Instituto Universitario de Microgravedad ���Ignacio Da Riva��� (IDR/UPM) of the Universidad Polit��cnica de Madrid. Application of Concurrent Engineering (CE) methodology in the framework of Multidisciplinary Design application (MDO) is one of the main objectives of the present work. In recent years, there has been continuing interest in the participation of university research groups in space technology studies by means of their own microsatellites. The involvement in such projects has some inherent challenges, such as limited budget and facilities. Also, due to the fact that the main objective of these projects is for educational purposes, usually there are uncertainties regarding their in orbit mission and scientific payloads at the early phases of the project. On the other hand, there are predetermined limitations for their mass and volume budgets owing to the fact that most of them are launched as an auxiliary payload in which the launch cost is reduced considerably. The satellite structure subsystem is the one which is most affected by the launcher constraints. This can affect different aspects, including dimensions, strength and frequency requirements. In the first part of this thesis, the main focus is on developing a structural design sizing tool containing not only the primary structures properties as variables but also the satellite system level variables such as payload mass budget and satellite total mass and dimensions. This approach enables the design team to obtain better insight into the design in an extended design envelope. The structural design sizing tool is based on the analytical structural design formulas and appropriate assumptions including both static and dynamic models of the satellite. A Genetic Algorithm (GA) is applied to the design space for both single and multiobejective optimizations. The result of the multiobjective optimization is a Pareto-optimal based on two objectives, minimum satellite total mass and maximum payload mass budget. On the other hand, the application of the microsatellites is of interest for their less cost and response time. The high need for the remote sensing applications is a strong driver of their popularity in space missions. The satellite remote sensing missions are essential for long term research around the condition of the earth resources and environment. In remote sensing missions there are tight interrelations between different requirements such as orbital altitude, revisit time, mission cycle life and spatial resolution. Also, all of these requirements can affect the whole design characteristics. During the last years application of the CE in the space missions has demonstrated a great advantage to reach the optimum design base lines considering both the performance and the cost of the project. A well-known example of CE application is ESA (European Space Agency) CDF (Concurrent Design Facility). It is clear that for the university-class microsatellite projects having or developing such a facility seems beyond the project capabilities. Nevertheless practicing CE at any scale can be beneficiary for the university-class microsatellite projects. In the second part of this thesis, the main focus is on developing a MDO framework applicable to the conceptual design phase of the remote sensing microsatellites. This approach enables the design team to evaluate the interaction between the different system design variables. The presented MDO framework contains not only the system level variables such as the satellite total mass and total power, but also the mission requirements like the spatial resolution and the revisit time. The microsatellite sizing process is divided into the three major design disciplines; a) orbit design, b) payload sizing and c) bus sizing. First, different mission parameters for a practical range of sun-synchronous orbits (SS-Os) are calculated. Then, according to the orbital parameters and a reference remote sensing instrument, mass and power of the payload are calculated. Satellite bus sizing is done based on mass and power calculation of the different subsystems using design estimation relationships. In the satellite bus sizing, the power subsystem design is realized by considering more detailed design variables including a mission scenario and different types of solar cells and batteries. The mission scenario is selected in order to obtain a coverage belt on the earth surface parallel to the earth equatorial after each revisit time. In order to evaluate the interrelations between the different variables inside the design space all the mentioned design disciplines are combined in a unified code. The integrated satellite system sizing tool developed in this section is considered as an application of the CE to the conceptual design of the remote sensing microsatellite projects. Finally, in order to apply the MDO methodology to the design problem, a basic MDO framework is adjusted to the developed satellite system design tool. Design optimization is done by means of a GA single objective algorithm with the objective function as minimizing the microsatellite total mass. According to the results of MDO application, there exist different optimum design points all with the minimum satellite total mass but with different mission variables. This output demonstrates the successful applicability of MDO approach for system engineering trade-off studies at the conceptual design phase of the design in such projects. The main conclusion of this thesis is that the classical design approach for the satellite design which usually starts with the mission and payload definition is not necessarily the best approach for all of the satellite projects. The university-class microsatellite is an example for such projects. Due to this fact an integrated satellite sizing tool including different design disciplines focusing on the structural subsystem and considering unknown payload is developed. According to the results the satellite total mass and available mass for the unknown payload are conflictive objectives. In order to find the Pareto-optimal a multiobjective GA optimization is conducted. Based on the optimization results it is concluded that selecting the satellite total mass in the range of 40-60 kg can be considered as an optimum approach for a university-class microsatellite project with unknown payload(s). Also, the CE methodology is applied to the remote sensing microsatellites conceptual design process. The results of CE application provide a clear understanding of the interaction between satellite system design requirements such as satellite total mass and power and the satellite mission variables such as revisit time and spatial resolution. The MDO application is done with the total mass minimization of a remote sensing satellite. The results from the MDO application clarify the unclear relationship between different system and mission design variables as well as the optimum design base lines according to the selected objective during the initial design phases.

Whole-genome genotyping of grape using a panel of microsatellite

The use of microsatellite markers in large-scale genetic studies is limited by its low throughput and high cost and labor requirements. Here, we provide a panel of 45 multiplex PCRs for fast and cost-efficient genome-wide fluorescence-based microsatellite analysis in grapevine. The developed multiplex PCRs panel (with up to 15-plex) enables the scoring of 270 loci covering all the grapevine genome (9 to 20 loci/chromosome) using only 45 PCRs and sequencer runs. The 45 multiplex PCRs were validated using a diverse grapevine collection of 207 accessions, selected to represent most of the cultivated Vitis vinifera genetic diversity. Particular attention was paid to quality control throughout the whole process (assay replication, null allele detection, ease of scoring). Genetic diversity summary statistics and features of electrophoretic profiles for each studied marker are provided, as are the genotypes of 25 common cultivars that could be used as references in other studies.

Analytical models for the power subsystem and the attitude control subsystem of a microsatellite

El trabajo realizado en la presente tesis doctoral se debe considerar parte del proyecto UPMSat-2, que se enmarca dentro del ��mbito de la tecnolog��a aeroespacial. El UPMSat-2 es un microsat��lite (de bajo coste y peque��o tama��o) dise��ado, construido, probado e integrado por la Universidad Polit��cnica de Madrid (Espa��a), para fines de demostraci��n tecnol��gica y educaci��n. El objetivo de la presente tesis doctoral es presentar nuevos modelos anal��ticos para estudiar la interdependencia energ��tica entre los subsistemas de potencia y de control de actitud de un sat��lite. En primer lugar, se estudia la simulaci��n del subsistema de potencia de un microsat��lite, prestando especial atenci��n a la simulaci��n de la fuente de potencia, esto es, los paneles solares. En la tesis se presentan m��todos sencillos pero precisos para simular la producci��n de energ��a de los paneles en condiciones ambientales variables a trav��s de su circuito equivalente. Los m��todos propuestos para el c��lculo de los par��metros del circuito equivalente son expl��citos (o al menos, con las variables desacopladas), no iterativos y directos; no se necesitan iteraciones o valores iniciales para calcular los par��metros. La precisi��n de este m��todo se prueba y se compara con m��todos similares de la literatura disponible, demostrando una precisi��n similar para mayor simplicidad. En segundo lugar, se presenta la simulaci��n del subsistema de control de actitud de un microsat��lite, prestando especial atenci��n a la nueva ley de control propuesta. La tesis presenta un nuevo tipo de control magn��tico es aplicable a la ��rbita baja terrestre (LEO). La ley de control propuesta es capaz de ajustar la velocidad de rotaci��n del sat��lite alrededor de su eje principal de inercia m��ximo o m��nimo. Adem��s, en el caso de ��rbitas de alta inclinaci��n, la ley de control favorece la alineaci��n del eje de rotaci��n con la direcci��n normal al plano orbital. El algoritmo de control propuesto es simple, s��lo se requieren magnetopares como actuadores; s��lo se requieren magnet��metros como sensores; no hace falta estimar la velocidad angular; no incluye un modelo de campo magn��tico de la Tierra; no tiene por qu�� ser externamente activado con informaci��n sobre las caracter��sticas orbitales y permite el rearme autom��tico despu��s de un apagado total del subsistema de control de actitud. La viabilidad te��rica de la citada ley de control se demuestra a trav��s de an��lisis de Monte Carlo. Por ��ltimo, en t��rminos de producci��n de energ��a, se demuestra que la actitud propuesto (en eje principal perpendicular al plano de la ��rbita, y el sat��lite que gira alrededor de ella con una velocidad controlada) es muy adecuado para la misi��n UPMSat-2, ya que permite una ��rea superior de los paneles apuntando hacia el sol cuando se compara con otras actitudes estudiadas. En comparaci��n con el control de actitud anterior propuesto para el UPMSat-2 resulta en un incremento de 25% en la potencia disponible. Adem��s, la actitud propuesto mostr�� mejoras significativas, en comparaci��n con otros, en t��rminos de control t��rmico, como la tasa de rotaci��n angular por sat��lite puede seleccionarse para conseguir una homogeneizaci��n de la temperatura m��s alta que apunta sat��lite y la antena. ABSTRACT The work carried out in the present doctoral dissertation should be considered part of the UPMSat-2 project, falling within the scope of the aerospace technology. The UPMSat-2 is a microsatellite (low cost and small size) designed, constructed integrated and tested for educational and technology demonstration purposes at the Universidad Polit��cnica de Madrid (Spain). The aim of the present doctoral dissertation is to present new analytical models to study the energy interdependence between the power and the attitude control subsystems of a satellite. First, the simulation of the power subsystem of a microsatellite is studied, paying particular attention to the simulation of the power supply, i.e. the solar panels. Simple but accurate methods for simulate the power production under variable ambient conditions using its equivalent circuit are presented. The proposed methods for calculate the equivalent circuit parameters are explicit (or at least, with decoupled variables), non-iterative and straight forward; no iterations or initial values for the parameters are needed. The accuracy of this method is tested and compared with similar methods from the available literature demonstrating similar precision but higher simplicity. Second, the simulation of the control subsystem of a microsatellite is presented, paying particular attention to the new control law proposed. A new type of magnetic control applied to Low Earth Orbit (LEO) satellites has been presented. The proposed control law is able to set the satellite rotation speed around its maximum or minimum inertia principal axis. Besides, the proposed control law favors the alignment of this axis with the normal direction to the orbital plane for high inclination orbits. The proposed control algorithm is simples, only magnetorquers are required as actuators; only magnetometers are required as sensors; no estimation of the angular velocity is needed; it does not include an in-orbit Earth magnetic field model; it does not need to be externally activated with information about the orbital characteristics and it allows automatic reset after a total shutdown of attitude control subsystem. The theoretical viability of the control law is demonstrated through Monte Carlo analysis. Finally, in terms of power production, it is demonstrated that the proposed attitude (on principal axis perpendicular to the orbit plane, and the satellite rotating around it with a controlled rate) is quite suitable for the UPMSat-2 mission, as it allows a higher area of the panels pointing towards the sun when compared to other studied attitudes. Compared with the previous attitude control proposed for the UPMSat-2 it results in a 25% increment in available power. Besides, the proposed attitude showed significant improvements, when compared to others, in terms of thermal control, as the satellite angular rotation rate can be selected to achieve a higher temperature homogenization of the satellite and antenna pointing.

An��lisis de algoritmos para posicionamiento de aeronaves mediante respuestas SSR utilizando recepci��n omnidireccional

El transporte a��reo constituye un sector estrat��gico para el crecimiento econ��mico de cualquier pa��s. El sistema de gesti��n de tr��fico a��reo ATM tiene como objetivo el movimiento seguro y eficiente de las aeronaves dentro del espacio a��reo y de los aeropuertos, siendo la seguridad, en la fase t��ctica, gestionada por el servicio de control de la circulaci��n a��rea. Mediante los procesos de control el tr��fico a��reo es vigilado a trav��s de sensores, regulado y guiado de forma organizada y segura. Es precisamente sobre la vigilancia donde se enfoca el contenido de la tesis, en el desarrollo de nuevos conceptos que proporcionen informaci��n de vigilancia de ���bajo coste��� basados en las se��ales existentes proporcionadas por la infraestructura actual de radar secundario y por los sistemas de posicionamiento basados en sat��lite que utiliza la ADS-B. El conocimiento y acceso en tiempo real a las trayectorias de las aeronaves es un elemento de valor a��adido no s��lo para la provisi��n de los servicios de control de tr��nsito a��reo, sino para todos los actores del transporte a��reo o de la investigaci��n, siendo uno de los elementos clave en el concepto operacional de los dos grandes proyectos tecnol��gicos, SESAR en Europa y NextGen en EE.UU.. En las ��ltimas d��cadas el control de la circulaci��n a��rea en espacios a��reos de media y alta densidad de tr��fico se ha basado en tecnolog��as complejas que requieren importantes infraestructuras como son el radar primario de vigilancia (PSR) y el radar secundario de vigilancia (SSR). La filosof��a de los programas SESAR y NextGen siguiendo las directrices de la OACI es la de alejarse de las tecnolog��as basadas en tierra para evolucionar hacia nuevas tecnolog��as m��s din��micas basadas en sat��lite como la ADS-B. Pero hasta que la implementaci��n y operaci��n de la ADS-B sea completa, existir�� un per��odo de transici��n que implica la coexistencia de aeronaves equipadas o no con ADS-B. El objetivo de la presente Tesis es determinar las metodolog��as y algoritmos m��s adecuados para poder hibridar las dos tecnolog��as descritas anteriormente, utilizando para ello un receptor de bajo coste con antena est��tica omnidireccional, que analice todas las se��ales presentes en el canal que comparten el SSR y ADS-B. Mediante esta hibridaci��n se podr�� obtener la posici��n de cualquier aeronave que transmita respuestas a interrogaciones SSR, en cualquiera de sus modos de trabajo, o directamente mensajes de posici��n ADS-B. Para desarrollar los algoritmos propuestos, adem��s del hardware correspondiente, se han utilizado las aplicaciones LabVIEW para funciones de adquisici��n de datos reales, y el software MATLAB�� para el desarrollo de algoritmos y an��lisis de datos. La validaci��n de resultados se ha realizado mediante los propios mensajes de posici��n ADS-B y a trav��s de las trazas radar proporcionadas por la entidad p��blica empresarial ENAIRE. La t��cnica desarrollada es aut��noma, y no ha requerido de ninguna otra entrada que no sea la recepci��n omnidireccional de las se��ales. Sin embargo para la validaci��n de resultados se ha utilizado informaci��n p��blica de las ubicaciones de la red de estaciones SSR desplegadas sobre territorio espa��ol y portugu��s y trazas radar. Los resultados obtenidos demuestran, que con t��cnicas basadas en superficies de situaci��n definidas por los tiempos de llegada de las respuestas, es posible determinar con una precisi��n aceptable la posici��n de las estaciones SSR y la posici��n de cualquier aeronave que responda mediante el Modo A a ��stas. ABSTRACT Air transport is a strategic sector for the economic growth of any country. The air traffic management system (ATM) aims at the safe and efficient movement of aircraft while operating within the airspace and airports, where safety, in the tactical phase, is managed by the air traffic control services. Through the air traffic control processes, aircraft are monitored by sensors, regulated and guided in an organized and safe manner. It is precisely on surveillance where this thesis is focused, developing new concepts that provide a 'low cost' surveillance information based on existing signals provided by currently secondary radar infrastructure and satellite-based positioning systems used by ADS-B. Having a deeper knowledge and a real-time access to the trajectories of the aircraft, is an element of added value not only for the provision of air traffic control services, but also for all air transport or research actors. This is one of the key elements in the operational concept proposed by the two large scale existing technological projects, SESAR in Europe and NextGen in the US. In recent decades, air traffic control in medium and high traffic density areas has been based on complex technologies requiring major infrastructures, such as the primary surveillance radar (PSR) and secondary surveillance radar (SSR). The philosophy of SESAR and NextGen programs, both following the guidelines of ICAO, is to move away from land-based technologies and evolving into some new and more dynamic satellite-based technologies such as ADS-B. Nevertheless, until the ADS-B implementation and operation is fully achieved, there will be a transitional period where aircraft with and without ADS-B equipment will have to coexist. The main objective of this thesis is to determine those methodologies and algorithms which are considered more appropriate to hybridize those two technologies, by using a low cost omnidirectional receiver, which analyzes all signals on the SSR and ADS-B shared channel. Through this hybridization, it is possible to obtain the position of any aircraft answering the SSR interrogations, in any of its modes of operation, or through the emission of ADS-B messages. To develop the proposed algorithms, LabVIEW application has been used for real-time data acquisition, as well as MATLAB software for algorithm development and data analysis, together with the corresponding hardware. The validation of results was performed using the ADS-B position messages and radar tracks provided by the Public Corporate Entity ENAIRE The developed technique is autonomous, and it does not require any other input other than the omnidirectional signal reception. However, for the validation of results, not only radar records have been used, but also public information regarding the position of SSR stations spread throughout the Spanish and Portuguese territory. The results show that using techniques based in the definition of positioning surfaces defined by the responses��� times of arrival, it is possible to determine with an acceptable level of accuracy both the position of the SSR stations as well as the position of any aircraft which transmits Mode A responses.

Caracterizaci��n de poblaciones de vid silvestre de la Pen��nsula Ib��rica

La vid silvestre se considera como el ancestro aut��ctono de las vides cultivadas y una enorme reserva gen��tica en peligro de extinci��n. La prospecci��n llevada a cabo entre 2003 y 2004 permiti�� catalogar 51 localizaciones de vides silvestres espa��olas, la mayor��a de ellas ubicadas en riberas de r��os. Estos ejemplares se incluyeron en el Banco de Germoplasma de la Finca "El Enc��n" (BGVCAM - Alcal�� de Henares, Madrid, Espa��a). En primer lugar, se caracteriz�� la cantidad y la distribuci��n de su diversidad gen��tica utilizando 25 loci empleando microsat��lites nucleares (SSR). Hemos analizado tambi��n la posible coexistencia en el h��bitat natural de vides silvestres con vides cultivadas naturalizadas y portainjertos. De este modo, los an��lisis fenot��picos y gen��ticos identificaron el 19% de las muestras recogidas como derivadas de genotipos cultivados, siendo, o bien vides cultivadas naturalizadas o genotipos h��bridos derivados de cruces espont��neos entre vides silvestres y cultivadas. La diversidad gen��tica de las poblaciones de vides silvestres fue similar a la observada en el grupo de las cultivadas. El an��lisis molecular mostr�� que el germoplasma de cultivadas y silvestres es gen��ticamente divergente con bajo nivel de introgresi��n. Se ha identificado cuatro grupos gen��ticos, con dos de ellos fundamentalmente representados por los genotipos de vides cultivadas y dos por las accesiones silvestres. El an��lisis de los v��nculos gen��ticos entre las vides silvestres y cultivadas podr��a sugerir una contribuci��n gen��tica de las accesiones silvestres espa��olas a las actuales variedades occidentales. En segundo lugar, se realiz�� un profundo estudio morfol��gico "ex situ " y se contrastaron con los resultados de la caracterizaci��n realizada en 182 variedades comerciales espa��olas de la misma colecci��n. Todos los individuos silvestres mostraron diferencias morfol��gicas con Vitis vinifera L subsp. vinifera, pero no se encontraron diferencias significativas dentro Vitis vinifera L. subsp. sylvestris, ni por localizaci��n geogr��fica ni por sexo. Los resultados de este estudio describen las principales caracter��sticas morfol��gicas de las vides silvestres espa��olas y sus rasgos diferenciales con su pariente cultivada. Por ��ltimo, se analiz�� la composici��n antoci��nica presente en 21 accesiones de vides silvestres de la Pen��nsula Ib��rica conservadas en el BGVCAM de la Finca "El Enc��n" y seleccionadas bas��ndose en diferencias ampelogr��ficas y caracterizaci��n molecular. La concentraci��n de antocianinas es similar la encontrad en vides cultivadas con destino a la vinificaci��n. Las accesiones estudiadas mostraron una variabilidad considerable en su perfil antoci��nico y fue posible distinguir varios grupos. Sin embargo, la presencia de material silvestre con perfiles antoci��nicos poco comunes o inexistentes en variedades espa��olas, sugiere que la variabilidad gen��tica relacionada con antocianinas en poblaciones espa��olas de vides silvestres podr��a ser m��s alta que la de variedades cultivadas com��nmente consideradas de origen espa��ol. ABSTRACT The wild grapevine is considered an autochthonous relative of cultivated vines and a huge gene pool endangered in Europe. Prospecting carried out between 2003 and 2004 enabled to inventory 51 Spanish sites with wild grapevines, most of them located near rivers. These individuals were grafted in the collection of "El Enc��n" (BGVCAM - Alcal�� de Henares, Madrid, Spain). Firstly, werw characterized the amount and distribution of their genetic diversity using 25 nuclear SSR loci. We have also analysed the possible coexistence in the natural habitat of wild grapevines with naturalized grapevine cultivars and rootstocks. In this way, phenotypic and genetic analyses identified 19% of the collected samples as derived from cultivated genotypes, being either naturalized cultivars or hybrid genotypes derived from spontaneous crosses between wild and cultivated grapevines. The genetic diversity of wild grapevine populations was similar than that observed in the cultivated group. The molecular analysis showed that cultivated germplasm and wild germplasm are genetically divergent with low level of introgression. We identified four genetic groups, with two of them fundamentally represented among cultivated genotypes and two among wild accessions. The analyses of genetic relationships between wild and cultivated grapevines could suggest a genetic contribution of wild accessions from Spain to current Western cultivars. Secondly, a morphological study was done "ex situ" and were compared with data from 182 Spanish commercial cultivars grown in the same collection. All wild individuals showed morphological differences with Vitis vinifera L. ssp. vinifera but no significant differences were found within Vitis vinifera L subsp. sylvestris neither by geographic origin nor by sex. A pattern with the main characteristics of Spanish wild grapevines is suggested. Ultimately, were investigated the anthocyanin composition of 21 mostly Spanish wild grapevine accessions preserved at BGVCAM "El Enc��n" and selected in consideration of observed ampelographic differences and molecular characterization. Total anthocyanin concentration was similar to that found in winegrape cultivars. The accessions studied showed considerable variability in their anthocyanin fingerprints and it was possible to distinguish several groups, similar to previous reports on the anthocyanin fingerprint of winegrapes. The anthocyanin composition of wild grapevine accessions was similar to that of cultivated grapes. Nevertheless, the presence of wild accessions with anthocyanin fingerprints uncommon or nonexistent in Spanish cultivated varieties suggests that the genetic variability related to anthocyanins in Spanish wild grapevine populations may be higher than that of cultivated varieties commonly considered of Spanish origin.