Molecular tools to improve chestnut management: El Bierzo as a case study

The European chestnut (Castanea sativa Mill.) is a multipurpose species that has been widely cultivated around the Mediterranean basin since ancient times. New varieties were brought to the Iberian Peninsula during the Roman Empire, which coexist since then with native populations that survived the last glaciation. The relevance of chestnut cultivation has being steadily growing since the Middle Ages, until the rural decline of the past century put a stop to this trend. Forest fires and diseases were also major factors. Chestnut cultivation is gaining momentum again due to its economic (wood, fruits) and ecologic relevance, and represents currently an important asset in many rural areas of Europe. In this Thesis we apply different molecular tools to help improve current management strategies. For this study we have chosen El Bierzo (Castile and Leon, NW Spain), which has a centenary tradition of chestnut cultivation and management, and also presents several unique features from a genetic perspective (next paragraph). Moreover, its nuts are widely appreciated in Spain and abroad for their organoleptic properties. We have focused our experimental work on two major problems faced by breeders and the industry: the lack of a fine-grained genetic characterization and the need for new strategies to control blight disease. To characterize with sufficient detail the genetic diversity and structure of El Bierzo orchards, we analyzed DNA from 169 trees grafted for nut production covering the entire region. We also analyzed 62 nuts from all traditional varieties. El Bierzo constitutes an outstanding scenario to study chestnut genetics and the influence of human management because: (i) it is located at one extreme of the distribution area; (ii) it is a major glacial refuge for the native species; (iii) it has a long tradition of human management (since Roman times, at least); and (iv) its geographical setting ensures an unusual degree of genetic isolation. Thirteen microsatellite markers provided enough informativeness and discrimination power to genotype at the individual level. Together with an unexpected level of genetic variability, we found evidence of genetic structure, with three major gene pools giving rise to the current population. High levels of genetic differentiation between groups supported this organization. Interestingly, genetic structure does not match with spatial boundaries, suggesting that the exchange of material and cultivation practices have strongly influenced natural gene flow. The microsatellite markers selected for this study were also used to classify a set of 62 samples belonging to all traditional varieties. We identified several cases of synonymies and homonymies, evidencing the need to substitute traditional classification systems with new tools for genetic profiling. Management and conservation strategies should also benefit from these tools. The avenue of high-throughput sequencing technologies, combined with the development of bioinformatics tools, have paved the way to study transcriptomes without the need for a reference genome. We took advantage of RNA sequencing and de novo assembly tools to determine the transcriptional landscape of chestnut in response to blight disease. In addition, we have selected a set of candidate genes with high potential for developing resistant varieties via genetic engineering. Our results evidenced a deep transcriptional reprogramming upon fungal infection. The plant hormones ET and JA appear to orchestrate the defensive response. Interestingly, our results also suggest a role for auxins in modulating such response. Many transcription factors were identified in this work that interact with promoters of genes involved in disease resistance. Among these genes, we have conducted a functional characterization of a two major thaumatin-like proteins (TLP) that belongs to the PR5 family. Two genes encoding chestnut cotyledon TLPs have been previously characterized, termed CsTL1 and CsTL2. We substantiate here their protective role against blight disease for the first time, including in silico, in vitro and in vivo evidence. The synergy between TLPs and other antifungal proteins, particularly endo-p-1,3-glucanases, bolsters their interest for future control strategies based on biotechnological approaches.

Desarrollo del teclado virtual “Mokey” basado en gestos para personas con movilidad reducida: capa de sistema

La Realidad Aumentada forma parte de múltiples proyectos de investigación desde hace varios años. La unión de la información del mundo real y la información digital ofrece un sinfín de posibilidades. Las más conocidas van orientadas a los juegos pero, gracias a ello, también se pueden implementar Interfaces Naturales. En otras palabras, conseguir que el usuario maneje un dispositivo electrónico con sus propias acciones: movimiento corporal, expresiones faciales, etc. El presente proyecto muestra el desarrollo de la capa de sistema de una Interfaz Natural, Mokey, que permite la simulación de un teclado mediante movimientos corporales del usuario. Con esto, se consigue que cualquier aplicación de un ordenador que requiera el uso de un teclado, pueda ser usada con movimientos corporales, aunque en el momento de su creación no fuese diseñada para ello. La capa de usuario de Mokey es tratada en el proyecto realizado por Carlos Lázaro Basanta. El principal objetivo de Mokey es facilitar el acceso de una tecnología tan presente en la vida de las personas como es el ordenador a los sectores de la población que tienen alguna discapacidad motora o movilidad reducida. Ya que vivimos en una sociedad tan informatizada, es esencial que, si se quiere hablar de inclusión social, se permita el acceso de la actual tecnología a esta parte de la población y no crear nuevas herramientas exclusivas para ellos, que generarían una situación de discriminación, aunque esta no sea intencionada. Debido a esto, es esencial que el diseño de Mokey sea simple e intuitivo, y al mismo tiempo que esté dotado de la suficiente versatilidad, para que el mayor número de personas discapacitadas puedan encontrar una configuración óptima para ellos. En el presente documento, tras exponer las motivaciones de este proyecto, se va a hacer un análisis detallado del estado del arte, tanto de la tecnología directamente implicada, como de otros proyectos similares. Se va prestar especial atención a la cámara Microsoft Kinect, ya que es el hardware que permite a Mokey detectar la captación de movimiento. Tras esto, se va a proceder a una explicación detallada de la Interfaz Natural desarrollada. Se va a prestar especial atención a todos aquellos algoritmos que han sido implementados para la detección del movimiento, así como para la simulación del teclado. Finalmente, se va realizar un análisis exhaustivo del funcionamiento de Mokey con otras aplicaciones. Se va a someter a una batería de pruebas muy amplia que permita determinar su rendimiento en las situaciones más comunes. Del mismo modo, se someterá a otra batería de pruebas destinada a definir su compatibilidad con los diferentes tipos de programas existentes en el mercado. Para una mayor precisión a la hora de analizar los datos, se va a proceder a comparar Mokey con otra herramienta similar, FAAST, pudiendo observar de esta forma las ventajas que tiene una aplicación especialmente pensada para gente discapacitada sobre otra que no tenía este fin. ABSTRACT. During the last few years, Augmented Reality has been an important part of several research projects, as the combination of the real world and the digital information offers a whole new set of possibilities. Among them, one of the most well-known possibilities are related to games by implementing Natural Interfaces, which main objective is to enable the user to handle an electronic device with their own actions, such as corporal movements, facial expressions… The present project shows the development of Mokey, a Natural Interface that simulates a keyboard by user’s corporal movements. Hence, any application that requires the use of a keyboard can be handled with this Natural Interface, even if the application was not designed in that way at the beginning. The main objective of Mokey is to simplify the use of the computer for those people that are handicapped or have some kind of reduced mobility. As our society has been almost completely digitalized, this kind of interfaces are essential to avoid social exclusion and discrimination, even when it is not intentional. Thus, some of the most important requirements of Mokey are its simplicity to use, as well as its versatility. In that way, the number of people that can find an optimal configuration for their particular condition will grow exponentially. After stating the motivations of this project, the present document will provide a detailed state of the art of both the technologies applied and other similar projects, highlighting the Microsoft Kinect camera, as this hardware allows Mokey to detect movements. After that, the document will describe the Natural Interface that has been developed, paying special attention to the algorithms that have been implemented to detect movements and synchronize the keyboard. Finally, the document will provide an exhaustive analysis of Mokey’s functioning with other applications by checking its behavior with a wide set of tests, so as to determine its performance in the most common situations. Likewise, the interface will be checked against another set of tests that will define its compatibility with different softwares that already exist on the market. In order to have better accuracy while analyzing the data, Mokey’s interface will be compared with a similar tool, FAAST, so as to highlight the advantages of designing an application that is specially thought for disabled people.

Discrimination of AE Signals From Friction and Concrete Cracking in a Reinforced Concrete Frame Subjected to Seismic Trainings

This paper shows the preliminary results of the development and application of a procedure to filter the Acoustic Emission (AE) signals to distinguish between AE signals coming from friction and AE signals coming from concrete cracking. These signals were recorded during the trainings of an experiment carried out on a reinforced concrete frame subjected to dynamic loadings with the shaking table of the University of Granada (Spain). Discrimination between friction and cracking AE signals is the base to develop a successful procedure and damage index based on AE testing for health monitoring of RC structures subjected to earthquakes.