Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.

Genetic redundancy among durum wheat accessions as assessed with SSRs and endosperm proteins

Reducing duplication in ex-situ collections is complicated and requires good quality genetic markers. This study was conducted to assess the value of endosperm proteins and SSRs for validation of potential duplicates and monitoring intra-accession variability. Fifty durum wheat (Triticum turgidum ssp. durum) accessions grouped in 23 potential duplicates, and previously characterised for 30 agro-morphological traits, were analysed for gliadin and high molecular weight glutenin (HMWG) subunit alleles, total protein, and 24 SSRs, covering a wide genome area. Similarity and dissimilarity matrices were generated based on protein and SSRs alleles. For heterogeneous accessions at gliadins the percent pattern homology (PH) between gliadin patterns and the Nei’s coefficient of genetic identity (I) were computed. Eighteen duplicates identical for proteins showed none or less than 3 unshared SSRs alleles. For heterogeneous accessions PH and I values lower than 80 identified clearly off-types with more than 3 SSRs unshared. Only those biotypes differing in no more than one protein-coding locus were confirmed with SSRs. A good concordance among proteins, morphological traits, and SSR were detected. However, the discrepancy in similarity detected in some cases showed that it is advisable to evaluate redundancy through distinct approaches. The analysis in proteins together with SSRs data are very useful to identify duplicates, biotypes, close related genotypes, and contaminations

Uso de marcadores moleculares para la racionalización y evaluación de la calidad en las colecciones de Recursos Fitogenéticos de trigo

Esta tesis tiene dos objetivos generales: el primero, analizar el uso de proteínas del endospermo y SSRs para la racionalización de las colecciones de trigo, y el segundo, estudiar la influencia de las proteínas del endospermo, del año de cultivo y del abonado nitrogenado en la calidad en un grupo de variedades locales españolas. Dentro del primer objetivo, se estudió la diversidad genética de la colección de Triticum monococcum L. (escaña menor), y de una muestra de la colección de Triticum turgidum L. (trigo duro) del CRF-INIA, con 2 y 6 loci de gliadinas, y 6 y 24 SSRs, para la escaña menor y el trigo duro, respectivamente. Ambas colecciones presentaron una gran diversidad genética, con una gran diferenciación entre las variedades y pequeña dentro de ellas. Los loci de gliadinas mostraron una gran variabilidad, siendo los loci Gli-2 los más útiles para distinguir variedades. En la escaña menor, las gliadinas presentaron mayor poder de discriminación que los SSRs; aunque en trigo duro los SSRs identificaron más genotipos. El número de alelos encontrado fue alto; 24 y 38 en gliadinas, y 29 y 203 en SSRs, en escaña menor y trigo duro, respectivamente. En trigo duro, se identificaron 17 alelos nuevos de gliadinas lo que demuestra que el germoplasma español es muy singular. En ambas especies, se detectaron asociaciones entre la variación alélica en prolaminas y el origen geográfico y filogenético de las variedades. La utilidad de las proteínas (6 loci de gliadinas, 2 loci de gluteninas y proteína total) y de los SSRs (24 loci) para verificar duplicados, y analizar la variabilidad intraaccesión, se estudió 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 proteína total, y diferencias en menos de tres loci de SSRs. El mismo resultado se obtuvo para los biotipos de la colección de T. monococcum. Sin embargo, las discrepancias observadas en algunos casos entre proteínas y SSRs demostraron la utilidad del uso conjunto de ambos tipos de marcadores. Tanto las proteínas como los SSRs mostraron gran concordancia con los caracteres agro-morfológicos, especialmente cuando las diferencias entre los genotipos eran grandes. Sin embargo, los caracteres agro-morfológicos fueron menos discriminantes que los marcadores moleculares. Para el segundo objetivo de la tesis, se analizó la variación alélica 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 incluía variedades locales de todas las provincias españolas donde se ha cultivado tradicionalmente el trigo duro. Todos los loci, excepto el Glu-B2, mostraron gran variabilidad genética, siendo los Glu-3 los más polimórficos. En total, se identificaron 65 alelos, de los que 29 eran nuevos, que representan una fuente importante de variabilidad genética para la mejora de la calidad. Se detectaron diferencias en la composición 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 común en la convar. turgidum, mientras que el Glu-B3a - Gli-B1c, asociado con mejor calidad, fue más frecuente en la convar. durum. En la convar. turgidum, se observó 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 proteína, P, y test de sedimentación, SDSS) con dos dosis de abonado nitrogenado (N), y en dos años diferentes. No se detectaron interacciones Variedad × Año, ni Variedad × N en la calidad. Para la P, los efectos ambientales (año y N) fueron mayores que el efecto de la variedad, siendo, en general, mayor la P con dosis altas de N. La variedad influyó más en el test SDSS, que no se vio afectado por el año ni el N. El aumento del contenido en proteína no influyó significativamente sobre la fuerza del gluten estimada con el SDSS. Respecto a la influencia de las prolaminas en la fuerza del gluten, se confirmó la superioridad del Glu-B3a; aunque también se detectó una influencia alta y positiva de los alelos nuevos Glu-A3new-1, y Glu-B3new-6 y new-9. La no correlación entre el rendimiento (evaluado en un trabajo anterior) y la P, en las variedades adaptadas a bajo N, permitió seleccionar cuatro variedades locales con alto rendimiento y buena fuerza del gluten para producción 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.

Problemática estructural y de funcionamiento de la cadena de valor del jamón ibérico de bellota: el caso de Guijuelo en España

El sector porcino ibérico es un sector típicamente español. La imagen que asocia el consumidor a los productos ibéricos es la correspondiente a los productos elaborados a partir de cerdos ibéricos en explotaciones extensivas, (dehesas), y cebados en montanera. Sin embargo, según los datos publicados por el Ministerio de Agricultura, Alimentación y Medio Ambiente, más de un 70 por ciento de los cerdos ibéricos que se producen en España son criados en granjas intensivas. El sector porcino ibérico está en pleno proceso de cambio, tanto por las variaciones que se derivan de la crisis que sufre el sector, como por las modificaciones que se esperan en la norma de calidad, recientemente acordadas. En este entorno en el que existe una reducción importante de los márgenes comerciales en todos los eslabones de la cadena de valor del jamón ibérico de bellota, adquiere importancia el estudio de su estructura y funcionamiento, la determinación de los aspectos que mejoran la eficiencia económica a lo largo de la misma y su concordancia a lo largo de ella. Así mismo, un sector de los consumidores comienza a buscar productos de calidad, se interesa por productos con determinados etiquetados que garantizan la calidad o unas determinadas prácticas de producción y elaboración, como sucede con los productos ecológicos. Existen numerosos estudios sobre distintos aspectos del sector porcino ibérico, realizados por distintas instituciones, como universidades, instituciones privadas y proyectos europeos. En general, se centran en la mejora de la producción de los cerdos ibéricos o en la mejora de la elaboración de los jamones. La novedad de la investigación realizada en la presente Tesis Doctoral reside en el estudio de los aspectos que determinan la mayor eficiencia económica de la cadena de valor del jamón ibérico de bellota de la denominación de origen de Guijuelo, y en las características que aporta al jamón ibérico de bellota la producción ecológica frente a la convencional. Resumen Los objetivos planteados en esta investigación son la mejora de la cadena de valor del jamón ibérico de bellota de la denominación de origen de Guijuelo, mediante el estudio de la problemática estructural y de funcionamiento de la misma. Como objetivos específicos se proponen la mejora de la eficiencia económica en las relaciones comerciales entre los distintos eslabones de la cadena de valor, y el impulso de la oferta de los productos ecológicos de ibérico, mediante el estudio comparativo de las producciones convencionales y ecológicas. Para alcanzar estos objetivos la metodología utilizada es la cadena de valor agroalimentaria, utilizando como herramientas de análisis el método Delphi y la realización de un análisis DAFO del sector porcino ibérico. Las fuentes de información primarias utilizadas proceden por una parte de la colaboración realizada en el proyecto europeo Q-Porkchains, coordinado por el Dr. Jacques H. Trienekens, y por otra parte de los cuestionarios especialmente diseñados para el análisis Delphi realizado. Las fuentes de información secundarias proceden de artículos académicos publicados, de artículos de revistas especializadas en el sector y de informes y estadísticas publicados por el Ministerio de Agricultura, Alimentación y Medio Ambiente. Las conclusiones de la investigación son las siguientes. Las relaciones comerciales de la cadena de valor estudiada están gestionadas, ya que se otorga gran valor a las relaciones comerciales basadas en la confianza, tanto en el aprovisionamiento de productos como en la venta de los mismos en cada eslabón. Sin embargo, no se gestiona adecuadamente la gama de productos que se oferta a lo largo de la cadena de valor, puesto que los eslabones de la misma obtienen diferente eficiencia económica para el mismo producto. Por otra parte, aunque la denominación de origen de Guijuelo es una marca conocida por el consumidor, el esfuerzo por tener una marca colectiva de calidad no ha sido totalmente satisfactorio, en especial para el eslabón de la distribución. Por esta razón, es importante resaltar que es recomendable la creación de una marca propia que cuide la calidad con especial atención, modificando incluso las empresas integrantes de la cadena de valor si el objetivo de calidad no coincide. Es importante garantizar la calidad del producto al consumidor, mediante la trasmisión la información generada por los informes de inspección y certificación realizados por las entidades correspondientes en relación a la norma de calidad de los productos de cerdo ibérico y por el Consejo Regulador de Guijuelo. Se requiere un estudio detallado de los posibles nichos de mercado para llegar a los clientes que buscan un producto de calidad diferenciada como es el jamón ibérico de denominación de origen de Guijuelo. Dentro de los posibles clientes debe tenerse en consideración al colectivo de personas alérgicas, ya que no se encuentran con facilidad en el mercado productos cárnicos de cerdo libres de alérgenos. Por último, la innovación es muy importante en este sector, y pueden ofertarse nuevos productos que ayuden al acortamiento de la cadena de valor, como podría ser la carne fresca de cerdo ibérico de bellota. La producción ecológica de porcino ibérico, en la actualidad no es significativa. El manejo que se realiza de los cerdos ibéricos convencionales de bellota es muy similar al manejo que se realiza de los cerdos ecológicos, salvo en la alimentación que reciben los cerdos ibéricos antes del cebo en montanera y en los medicamentos permitidos en ambas producciones. La producción de porcino ecológico de bellota puede ser interesante para la exportación. Los jamones ibéricos de bellota dispondrían de un sello que garantiza un manejo tradicional –como se realiza en muchas explotaciones extensivas de cerdos ibéricos de bellota-, y que es apreciado por los consumidores de otros países, dispuestos a pagar por productos de calidad diferenciada. Este tipo de producción quizá podría solventar la limitación al ámbito nacional de la norma de calidad de los productos de cerdo ibérico. La falta de claridad y unanimidad en el sector sobre la calidad de los productos de ibéricos que se percibe en los cambios previstos en la legislación, y el interés del eslabón de la industria elaboradora de jamones en calidades no ligadas a producciones extensivas sino intensivas, por su mayor volumen de ventas y sus mejores rendimientos, mueven a los productores de “ibérico puro de bellota” a desmarcarse del sector “ibérico”, donde el peso del producto ibérico cruzado de pienso tiene mucho peso en las decisiones que se toman. The Iberian pork industry is a typically Spanish industry. The associated image by the consumer to these products corresponds to products made from Iberian pigs in extensive farms (meadows), and fattened in open range. However, according to data released by the Ministry of Agriculture, Food and Environment, more than 70 percent of the Iberian pigs produced in Spain are raised on factory farms. The Iberian pork industry is in process of change, both by modifications resulting from the crisis in the sector, and the expected ones in the quality standard, recently agreed. In this environment where there is a significant reduction in trade margins in all links of the value chain of Iberian ham, becomes important to study its structure and operation, identifying the aspects that improve economic efficiency along the value chain and their concordance along it. Likewise, part of the consumers begins searching for quality products, they are interested in certain products that guarantee some sort of quality or production and processing practices, like organic products. There are several studies on various aspects of the Iberian pork industry, made by different institutions, such as universities, private institutions and some European projects. In general, their goal is focused on improving the Iberian hog production or on improving the elaboration of hams. The novelty of the research conducted in this PhD thesis lies in the study of the aspects that determine the economic efficiency of the value chain of Iberian ham from Guijuelo designation of origin, and the comparison between the features that organic or conventional production of Iberian ham brings to it. The objectives propound in this research are the improvement of the value chain of Iberian ham from Guijuelo designation of origin, through the study of its structural and operational problematic. The proposed specific objectives are the improvement of the economic efficiency Resumen in trade relations between the different links in the value chain, and the promotion of the supply of Iberian organic pork products, through the comparative study of conventional and organic productions. In order to achieve these objectives, the used methodology is the agrifood value chain, using as analysis tools the Delphi method, and a SWOT analysis of the Iberian pork industry. The primary sources of information come partly from the collaboration in the European project Q-Porkchains, coordinated by Dr. Jacques Trienekens, and partly from the questionnaires specially designed for the Delphi analysis performed. The secondary sources come from published academic papers, specialized journal articles and reports published by the Ministry of Agriculture, Food and Environment. The research conclusions are as follows. Trade relations in the studied value chain are managed, as it is granted a great value to business relationships based on trust, both in the supply of products and in the sale in each link. However, the portfolio that is offered throughout the value chain is not properly managed, since the economic efficiency obtained for the same product is different in each link. Moreover, although the designation of origin of Guijuelo is a well-known brand, the effort made to acquire a collective quality trademark has not been entirely satisfactory, especially in the distribution link. Due to it, it is important to highlight that it is recommended to create a brand focused on quality, and indeed to change the firms integrating the vale chain, if the goal is not shared by all the companies. It is essential to ensure the quality of the product to the consumer, through the transmission of the available information related to the quality standard of Iberian pork products and of the Guijuelo Regulating Council. A detailed study of potential niche markets is required, in order to reach customers looking for a differentiated quality product as Iberian ham of Guijuelo designation of origin. It should be taken into consideration the group of allergy sufferers as potential customers, as they do not easily find pork products in the market free of allergens. Innovation is very important in this industry, and new products could be commercialized that help to shorten the value chain, as fresh acorn Iberian pork. Nowadays, the organic production of Iberian pork products is not significant. The operation is very similar in both conventional and organic production of Iberian pork products, except in the food received by Iberian hogs and the sort of medicinal treatments allowed in each production. The organic acorn Iberian pork production could be interesting for export. Acorn Iberian hams would have a well-known hallmark, which guarantees a traditional production, -as it is done in many extensive holdings of Iberian acorn-pigs-, and that is appreciated by consumers in other countries This niche of consumers could pay for differentiated quality products. Perhaps, this sort of production could solve the limitation to national scope of the quality standard of Iberian pork products. The lack of clarity and unanimity in the industry on the quality of Iberian pork products has been perceived in the agreed changes of the legislation as well as the interest of the processing industry in hams of quality categories which are not linked to extensive but intensive productions, due to their higher sales volume and better yields. This situation pushed "acorn pure Iberian " pigs farmers to split from the Iberian industry, where the importance of cross Iberian pig from intensive farms is growing so high that they can influence the negotiations and decisions taken inside and outside the industry.

Reducing the dimensionality effect in importance sampling simulations

The dimensionality effect is avoided by the use of sufficient statistics in event probability estimators realised by importance sampling. If the system function is not a sufficient statistic, an approach is proposed to reduce the dimensionality effect in the estimators. Simulation results of false-alarm probability estimations, applied to radar detection, confirm a clear concordance with the theoretical results

Analysis of air flow past and through the 2415-3S airfoil for an unmanned aerial vehicle with internal propulsion system

This paper deals with the prediction of velocity fields on the 2415-3S airfoil which will be used for an unmanned aerial vehicle with internal propulsion system and in this way analyze the air flow through an internal duct of the airfoil using computational fluid dynamics. The main objective is to evaluate the effect of the internal air flow past the airfoil and how this affects the aerodynamic performance by means of lift and drag forces. For this purpose, three different designs of the internal duct were studied; starting from the base 2415-3S airfoil developed in previous investigation, basing on the hypothesis of decreasing the flow separation produced when the propulsive airflow merges the external flow, and in this way obtaining the best configuration. For that purpose, an exhaustive study of the mesh sensitivity was performed. It was used a non-structured mesh since the computational domain is three-dimensional and complex. The selected mesh contains approximately 12.5 million elements. Both the computational domain and the numerical solution were made with commercial CAD and CFD software, respectively. Air, incompressible and steady was analyzed. The boundary conditions are in concordance with experimental setup in the AF 6109 wind tunnel. The k-e model is utilized to describe the turbulent flow process as followed in references. Results allowed obtaining velocity contours as well as lift and drag coefficients and also the location of separation and reattachment regions in some cases for zero degrees of angle of attack on the internal and external surfaces of the airfoil. Finally, the selection of the configuration with the best aerodynamic performance was made, selecting the option without curved baffles.

Numerical analysis of air flow past the 2415-3S airfoil for an unmanned aerial vehicle with internal propulsion system

This paper deals with the prediction of pressure and velocity fields on the 2415-3S airfoil which will be used for and unmanned aerial vehicle with internal propulsion system and in this way analyze the air flow through an internal duct of the airfoil using computational fluid dynamics. The main objective is to evaluate the effect of the internal air flow past the airfoil and how this affects the aerodynamic performance by means of lift and drag forces. For this purpose, three different designs of the internal duct were studied; starting from the base 2415-3S airfoil developed in previous investigation, basing on the hypothesis of decreasing the flow separation produced when the propulsive airflow merges the external flow, and in this way obtaining the best configuration. For that purpose, an exhaustive study of the mesh sensitivity was performed. It was used a non-structured mesh since the computational domain is tridimensional and complex. The selected mesh contains approximately 12.5 million elements. Both the computational domain and the numerical solution were made with commercial CAD and CFD software respectively. Air, incompressible and steady was analyzed. The boundary conditions are in concordance with experimental setup in the AF 6109 wind tunnel. The k-ε model is utilized to describe the turbulent flow process as followed in references. Results allowed obtaining pressure and velocity contours as well as lift and drag coefficients and also the location of separation and reattachment regions in some cases for zero degrees of angle of attack on the internal and external surfaces of the airfoil. Finally, the selection of the configuration with the best aerodynamic performance was made, selecting the option without curved baffles.