Molecular mechanisms of Strawberry Plant Defence against colletotrichum acutatum

This thesis is focused on strawberry molecular studies aimed by the strong economic impact and social staple that represents this crop. With an annual production of 500000 tons and an economic weigh of 650 million €, Spain is the third producing country in world (FAOSTAT Agriculture Data [http://faostat.fao.org/]). Important losses in strawberry yields occur due to diseases and pests. Although resistant cultivars are a priority of most strawberry breeding programs, completely resistant cultivars have not yet been reported, relying pathogen control on the excessive use of chemical products (mostly environmental contaminants). Despite of the immediate necessity on developing new strategies to improve resistance in this crop plant, molecular knowledge is still scarce and most components and mechanisms of the strawberry defense network remain unknown and poorly understood. The main aim of this work is to get molecular clues about how plant immunity is activated in strawberry to face pathogen attack. Understanding the molecular interplay between strawberry plant and microbes will successful contribute to identify candidate genes useful for developing biotechnological strategies and help breeding to increase resistance against specific pathogens. This work has been structured in eight chapters which contributed to mount and exploit new technical platforms to subsequently uncover the strawberry defense response at molecular level. Chapter I is an intense and comprehensive compendium of all available information on strawberry immunity, and contribute to the field by discussing, updating and compiling research focused on the molecular aspects and events of the strawberry defense mechanisms against pathogens, concluding that major progress in the physiology, genetics and molecular biology of strawberry, is still needed to fully uncover the logic of its elaborate plant innate immune system. In this context, the use of high-throughput technologies will provide large amount of molecular information related with defense response in strawberry. However, the lack on commercially-available tools focused on crop plants such as strawberry, aimed us to produce our private platforms to be exploited in strawberry-based studies, as it is described in Chapter II. Thus, the generation of a ESTs collection enriched on defense-related genes, which allow the identification of new components of strawberry immunity, and provides information on those biological...Esta tesis está enfocada en estudios moleculares de la planta de fresa animada por el importante impacto económico y estabilizador social que representa este cultivo. Con una producción anual de 500000 toneladas y una relevancia económica de 650 millones de euros, España es el tercer país productor en el mundo (FAOSTAT Agriculture Data [http://faostat.fao.org/]). La producción de fresa sufre importantes pérdidas como consecuencia de las enfermedades y las plagas. Aunque la obtención de cultivares resistentes son una prioridad para la mayoría de los programas de mejora, aún no se han obtenido cultivares completamente resistentes, por lo que el control de los patógenos ha recaído en el uso excesivo de productos químicos (mayoritariamente contaminantes ambientales). Aún con la urgente necesidad de desarrollar nuevas estrategias para mejorar la resistencia en este cultivo, el conocimiento a nivel molecular de la mayoría de los componentes y mecanismos de la respuesta de defensa de la planta de fresa permanecen desconocidos y difícilmente entendibles. El objetivo principal de este trabajo consiste en la obtención de pistas a nivel molecular a cerca de cómo la inmunidad de la planta de fresa es activada para enfrentarse al ataque de los patógenos. Conocer la interacción molecular entre la planta de fresa y los microbios contribuirá eficazmente a la identificación de genes candidatos útiles en el desarrollo de estrategias biotecnológicas y ayudará en los programas de mejora para incrementar la resistencia contra patógenos específicos. Este trabajo se ha estructurado en ocho capítulos que contribuyen en la tarea de montar y explotar nuevas plataformas técnicas para descubrir secuencialmente, a un nivel molecular, los distintos pasos en la respuesta de defensa desplegados por la planta de fresa. El Capítulo I es un amplio e intenso compendio de toda la información disponible sobre la inmunidad de la fresa, y contribuye a este campo con la discusión, actualización y compilación de toda la investigación enfocada en los aspectos moleculares de los mecanismos de la defensa de esta planta. Este capítulo concluye que aún es necesario un importante esfuerzo en los estudios de la fisiología, la genética y la biología molecular en fresa, para llegar a discernir por completo la lógica de su muy elaborado sistema de inmunidad innata. En este contexto, el uso de tecnologías de gran escala proporcionará una gran cantidad de información molecular relacionada con la respuesta de defensa en..

Influencia de las sustituciones isomórficas en los factores de estructura de hidroxisales de Co-Cu; Cu-Zn; Zn-Mg y Mg-Cu.

Depto. de Mineralogía y PetrologíaFac. de Ciencias GeológicasTRUEpu

A Comprehensive Study of the WRKY Transcription Factor Family in Strawberry

WRKY transcription factors play critical roles in plant growth and development or stress responses. Using up-to-date genomic data, a total of 64 and 257 WRKY genes have been identified in the diploid woodland strawberry, Fragaria vesca, and the more complex allo-octoploid commercial strawberry, Fragaria × ananassa cv. Camarosa, respectively. The completeness of the new genomes and annotations has enabled us to perform a more detailed evolutionary and functional study of the strawberry WRKY family members, particularly in the case of the cultivated hybrid, in which homoeologous and paralogous FaWRKY genes have been characterized. Analysis of the available expression profiles has revealed that many strawberry WRKY genes show preferential or tissue-specific expression. Furthermore, significant differential expression of several FaWRKY genes has been clearly detected in fruit receptacles and achenes during the ripening process and pathogen challenged, supporting a precise functional role of these strawberry genes in such processes. Further, an extensive analysis of predicted development, stress and hormone-responsive cis-acting elements in the strawberry WRKY family is shown. Our results provide a deeper and more comprehensive knowledge of the WRKY gene family in strawberry

Proteomic analysis of goat milk kefir: profiling the fermentation-time dependent protein digestion and identification of potential peptides with biological activity

Kefir is a fermented dairy product, associated to health benefits because of being a probiotic and due to the presence of molecules with biological activity. In this work, we have profiled the peptide composition of goat milk kefir at three different fermentation times using a peptidomics approach, in order to study changes in peptide concentrations and patterns of protein digestion throughout the fermentation time. We identified 2328 unique peptides corresponding to 22 protein annotations, with a maximum of peptides found after 24 h fermentation. We established different digestion patterns according to the nature of the proteins, and quantified the changes in the peptides appearing in all the fermentation times. We also identified 11 peptides that matched exactly to sequences with biological activity in databases, almost all of them belonging to caseins. This is the most comprehensive proteomic analysis of goat milk kefir to date

Ion Torrent and lllumina, two complementary RNA-seq platforms for constructing the holm oak (Quercus ilex) transcriptome

Transcriptome analysis is widely used in plant biology research to explore gene expression across a large variety of biological contexts such as those related to environmental stress and plant-pathogen interaction. Currently, next generation sequencing platforms are used to obtain a high amount of raw data to build the transcriptome of any plant. Here, we compare Illumina and Ion Torrent sequencing platforms for the construction and analysis of the holm oak (Quercus ilex) transcriptome. Genomic analysis of this forest tree species is a major challenge considering its recalcitrant character and the absence of previous molecular studies. In this study, Quercus ilex raw sequencing reads were obtained from Illumina and Ion Torrent and assembled by three different algorithms, MIRA, RAY and TRINITY. A hybrid transcriptome combining both sequencing technologies was also obtained in this study. The RAY-hybrid assembly generated the most complete transcriptome (1,116 complete sequences of which 1,085 were single copy) with a E90N50 of 1,122 bp. The MIRAIllumina and TRINITY-Ion Torrent assemblies annotated the highest number of total transcripts (62,628 and 74,058 respectively). MIRA-Ion Torrent showed the highest number of shared sequences (84.8%) with the oak transcriptome. All the assembled transcripts from the hybrid transcriptome were annotated with gene ontology grouping them in terms of biological processes, molecular functions and cellular components. In addition, an in silico proteomic analysis was carried out using the translated assemblies as databases. Those from Ion Torrent showed more proteins compared to the Illumina and hybrid assemblies. This new generated transcriptome represents a valuable tool to conduct differential gene expression studies in response to biotic and abiotic stresses and to assist and validate the ongoing Q. ilex whole genome sequencing